Novel method for chromatographic finger printing and standardization of single medicines and formulations

ABSTRACT

The present invention provides a method for the chromatographic fingerprinting, chemical and therapeutic standardization, bar-coding of the fingerprints and preparation of a data base for Enterprise Resource Planning (ERP) and Customer Relationship Management (CRM) machines and applications of medicines in general and traditional medicines in particular; this invention includes a software based instrumental method and a novel method of fingerprinting and standardization is proposed for the above purpose and the said method for the chromatographic finger printing which facilitates to correlate the traditional therapeutic standardization methods with the chemical properties of the medicines and humors and provides a rational basis to understand the methods used for the said purpose.

TECHNICAL FIELD

The present invention relates to a novel method of chromatographicfinger printing useful for chemical and therapeutic standardization.More particularly, the present invention relates to chromatographicfingerprinting of organic and organo-metallic molecules which haveUV-Visible radiation absorptive property present in a plant, animal orany other source capable of being used as a single or formulatedmedicine. The invention facilitates bar coding of one or manyconstituents present in the finger print images. It also facilitatecommercial utilization of the fingerprint database having all theinformation about the medicines, development and using ERP (EnterpriseResource Planning) and CRM (Customer Relationship Management) net workapplications like vending machines etc.

The present invention employs a novel method of the utilization of theContour and 3D chromatograms of an herbal medicine and formulationdeveloped under standardized experimental (chemical and instrumental)conditions proposed as a chromatographic finger print of the medicines.

In addition, the present invention relates to a computer-based methodfor the analysis of such chromatograms. This novel method is very muchused for an authentic identification of the chemical constituents of thesingle medicines and formulations.

BACKGROUND ART

History of Traditional Medicines in the World

The ancient man after many years of evolution started thinking about himand tried to understand the nature. He started living under a cover, thecaves and in-groups. The process of thought has encouraged him tounderstand the nature and its inter action with the living beings. Hestarted using the naturally available flora and fauna for his dailyneeds, in which he used the plant and animal material for his dietaryand health needs.

Under this process, he explored the properties of various components inthe world, like geological, astral and medicinal properties of variousbiological and plant materials. This started happening from the StoneAge on words. He went on discovering, standardizing and using naturallyavailable materials for his day-to-day living.

This took place in many places of the earth parallel in different partsof the world and it developed more in places where civilization hasdeveloped more based on the intellectuality. Therefore, the history ofmedicine has a direct relation with the history of civilization.

The evidences of an well-organized system of medicine in India weretraced in Harappa and Mohanzadaro (History of Medicine in India, DrPriya Vrit Sharma, Indian National Science Academy, New Delhi). In theIndus valley civilization, a system of medicine has prevailed, in whichdrugs of vegetable, animal and mineral origin were used. The Osadhisuktaof the Rigveda is the oldest document of the knowledge about plants andherbal medicines. Medicine in India owes much to the traditionalknowledge of Atharvaveda of which Ayurveda is said as an Upaveda. Alarge number of disease-syndrome relationships were defined anddescribed by Charaka and Susruta in their medical treatises ‘TheSamhitas’. The treatment was also prescribed in a systematic manner andon rational basis.

On the other hand, it was realized that the biological phenomena couldnot be universally explained by mechanical means as each individualvaries in his basic constitution i.e., Prakruthi that must be kept inmind while prescribing diet or drug to the patient. The binary conceptlike Prakriti-Purusha, Yin-Yang, Normal-Abnormal was seen in almost allphilosophies.

Diseases are manifestations of humoral imbalance, which have to betackled comprehensively on the psychosomatic basis. Health accordinglyis equilibrium maintained on physical, mental and spiritual levels.Thus, the Tri-dimensional definition of health as propounded by Susrtutais the ideal one, which has been reflected in the definition, adopted byWHO in modern times.

After going through the ancient literature it was found that, themedicines were standardized using their physico-chemical properties ofthe materials. The color, texture, odor and taste were used as a measureof the efficacy of any medicine. Even the shape of the medicines wasused to understand the medicinal properties of the medicines. A summaryof different philosophies and various factors used in the therapy isgiven in Tables 1-6 which are appearing in the later part of thedescription. FIG. 1 accompanying the specification gives a detailedinformation on the individual philosophies and concepts of IndianAyurveda and Chinese Traditional medicines (Medicine in China by H. M.Sais and A hand book of Chinese Healing Herbs by Daniel Reid, Simon &Schuster) in particular and other philosophies in general. Thetherapeutic efficacy of a medicine ultimately depends on the chemicalconstituents present in the medicines under use and it is the chemicalproperties of the constituents that are going to bring a required changein the chemical constitution of the living animal.

Many Ayurvedic (one kind of Indian system of medicine) scholars havedefined and classified the medicines based on color and theirtherapeutic efficacy. A brief summary is given in Table 7, which appearsin the later part of the description. Thus, the physico-chemicalproperties of the materials and man is taken in to consideration tounderstand the properties of them and use for the therapy to achieverequired therapeutic results. Tables 8-9 which are appearing in thelater part of the description gives the information about how thephysical properties (color) and chemical properties (taste) are used forunderstanding the therapeutic efficacy of medicines and their influenceon the physiology of the human body. One part of the present work alsouses the same methodology, but with instruments.

In general, the constituent molecules present in the drugs and foods canbe broadly classified in to three categories as polar, medium polar andthe non-polar molecules. The total polarity of the molecule depends onthe total Electrophlic and Nucleophilic moieties attached to themolecule along with the unsaturation of the molecules by theirconjugation. The living human body, animal body and plants will alsocontain the same type of molecules wherein different polar moleculeswill carry out different functions. Diseases were cured using themedicines of same polarity as that of the disease causing chemicalconstituents, i.e. the medicines which can create the disorder can curethe same disorder, as said by Dr Hanemann.

WHO Definition of Herbal Medicine:

WHO has defined herbal medicine as a “Finished, labeled medicinalproducts that contain as active ingredients aerial or underground partsof plants, or other plant material, or combinations thereof whether inthe crude state or as plant preparations. Plant material includesjuices, gums, fatty oils, essential oils, and any other substances ofthis nature. Herbal medicines may contain excipients in addition to theactive ingredients. Medicines containing plant material combined withchemically defined active substances, including chemically defined,isolated constituents of plants, is not considered to be herbalmedicines. Exceptionally, in some countries herbal medicines may alsocontain, by tradition, natural organic or inorganic active ingredientswhich are not of plant origin”.

The objective of these guidelines, therefore, is “to define basiccriteria for the evaluation of quality, safety, and efficacy of herbalmedicines and thereby to assist national regulatory authorities,scientific organizations, and manufacturers to undertake an assessmentof the documentation/submission/dossiers in respect of such products”.As a general rule in this assessment, traditional experience means thatlong-term use as well as the medical, historical and ethnologicalbackground of those products shall be taken into account. The definitionof long-term use may vary according to the country but should be atleast several decades. Therefore, the assessment should take intoaccount a description in the medical/pharmaceutical literature orsimilar sources, or a documentation of knowledge on the application ofan herbal medicine without a clearly defined time limitation. Marketingauthorizations of similar products should also be taken into account. Asper the report, the assessment of quality should be done for thefollowing parameters.

WHO guidelines were given for the finished product for which, theassessment of efficacy, activity, evidence required to supportindications and combination products. Many herbal remedies consist of acombination of several active ingredients, and as experience of the useof traditional remedies is often based on combination products,assessment should differentiate between old and new combinationproducts. Identical requirements for the assessment of old and newcombinations would result in inappropriate assessment of certaintraditional medicines. In the case of traditionally used combinationproducts, the documentation of traditional uses (such as classical textsof Ayurveda, TraditionaL Chinese medicine, Unani and Siddha) andexperience may serve as evidence of efficacy.

An explanation of new combination of well known substances includingeffective dose ranges and compatibility should be required in additionto the documentation of traditional knowledge of each single ingredient.Each active ingredient must contribute to the efficacy of the medicine.Clinical studies may be required to justify the efficacy of a newingredient and its positive effect on the total combination.

In the report, it was also mentioned that the manufacturing procedureand formula, including the amount of excipients, should be described indetail. A finished product specification should be defined. A method ofidentification and where possible, quantification of the plant materialin the finished product should also be defined. If the identification ofan active principle is not possible, it should be sufficient to identifya characteristic substance or mixture of substances (e.g.,“chromatographic fingerprint”) to ensure consistent quality of theproduct. The finished product should comply with general requirementsfor particular dosage forms.

For imported finished products, confirmation of the regulatory status inthe country of origin should be provided. The WHO Certification Schemeon the quality of pharmaceutical products moving in internationalcommerce should be applied. More details about stability, assessment ofsafety and utilization were given in the said WHO report.

The effective regulation of the quality of herbal medicines moving ininternational commerce also requires close liaison between nationalinstitutions that are able to keep under regular review all aspects ofproduction and use of herbal medicines. Also, to conduct or sponsorevaluative studies of their efficacy, toxicity, safety, acceptability,cost and relative value are compared with other drugs used in modernmedicine.

Hence, as mentioned above, there is a need for an authentic method ofquality control as proposed in this work. It is clearly mentioned thatthere is a need of a method for all the above purposes. The proposedanalytical will give answers for almost all of the needs describedabove.

Existing Methods of Standardization:

Before explaining the method of the invention of standardization, theexisting methods of standardization (chemical and therapeutic) andchromatographic finger printing are discussed below.

A. Prior Art on Chemical Standardization:

I) Traditional:

The great sage Charaka explained in his Charaka Samhita that “Theunderstanding of the totality of an entity does not arise from afragmentary knowledge of it” (Charaka Samhita VI. 4.5). This makes itclear that standardization and therapeutic efficacy of any medicine forwhich all the constituents are not taken into consideration is futile.

The qualitative and quantitative profile of a herbal medicine will varydue to many geological, ecological factors, time of collection, place ofcollection, age of collection and weather conditions at the time ofcollection and so on.

Traditional herbalists used to select a medicine based on theorganoleptic methods available at that time like color, texture, smelland taste by which they used to assess the chemical and therapeuticefficacy of a medicine.

These methods involve intrinsic knowledge and understanding of the interand intra therapeutic interactions of the medicines and bodyconstituents to cure diseases. This knowledge varies from individual toindividual and depends on the individual skill and ability. Practically,it will be difficult to provide a rational justification for anymechanism to explain, using this method. Hence, modern science usesinstruments for various purposes, which eliminates the individualfactors and facilitates reproducibility in data and information.

ii) Modern:

The therapeutic property of any food or drug will depend on its chemicaland physical status. Thus, understanding the chemical constituents usingtheir physico-chemical properties will help to understand thetherapeutic efficacy of the medicine.

The physico chemical properties of the medicines play a major role onthe therapeutic activity of the medicine. These properties of moleculescan be studied using two parameters, the polarity and conjugativeproperties. Polarity is a resultant electrochemical property due todifferent electron donating (nucleophilic) and electron accepting(electrophilic) moieties attached to the molecules along with theunsaturated double and triple bonds present in it. They will influencethe rate of activity or reactivity of a molecule in chemical andbiochemical reactions. A thorough estimation of the total polarity ofthe molecule will give the efficacy of a single or group of molecules asto how active they are chemically and therapeutically. Hence, anystandardization, which assesses the above properties, will be useful toknow their activity.

Along with the polarity, which relates mostly to the Electro-chemicalproperty of the molecules the physical structure of the molecule alsoplay an important role in the reactivity of the molecules. The more thenumber of active sites attached to the molecules, the more reactive theywill be. The more the molecule is conjugated (having alternative doublebond and triple bonds) the more it will be reactive chemically and sotherapeutically.

The second parameter that influences the activity of the molecule is thespatial arrangement of atoms in the same molecule, which differsstructurally. Due to this reason, the isomeric (geometric and chiral)molecules play an important role in the biological activity. Thisstereo-selective nature makes the molecules highly selective in theiractivity in the body where in a large number of biochemical pathwayswill be working parallel without cross interactions and interferences.Hence, the chemistry of chiral drugs has become very important. In otherterms, no key (chiral molecule) will open a different lock (receptor).

Plants usually prepare a plurality/combinatorial library of moleculeshaving the same basic mother structures and varying in the functionalgroups attached to it. For example, flavones, aurones and chalcones offlavoniods as they exist in nature and a single plant having such set ofmolecules will act like a multi drug.

Usually molecules having unsaturation and more conjugation absorbs theelectromagnetic radiation in the UV-Visible radiation (200-800 nm). Whenthe compounds interact with radiation they absorb at a specificwavelength (absorbance maxima) based on their chemical, conjugative andstructural properties. It is called the characteristic wavelength. Amolecule can have more than one-absorbance maxima based on itsstructural and functional properties. When a compound absorbs aparticular color from the wholesome of the white light it will expressthe resultant color of the other colors unabsorbed. Thus, the materialswill express different colors based on their chemical constituentsabsorbing various colors from white light and showing that their coloris due to the various functional groups attached to it. (Table No 10,which appears later part of the description, explains the same). Thesame is taken as a measure of the chemical and physical properties ofthe molecules in spectrophotometry.

For example, the red colored medicines absorbs at 500-600 nm range.Thus, all the red colored medicines will have a peak in this wavelengthrange having specific structure and activity. Hence, the color of themedicines is being used as a measure of its therapeutic and chemicalefficacy of the medicines. In ancient times, medicines were classifiedtherapeutically based on the color. The present method proves the same.The FIG. 2 which accompanying the specification depicts relation ofcolors with humors shows the effect of different colors on differentdiseases.

FIG. 3, which accompany the specification, the fingerprints of twoShilajit samples prepared by the method of the invention, shows thedifference in the chemical profile of both samples. Shilajit is acarbonaceous material that forms due to storage of vegetable and animalmatter inside the earth for many years. Mostly due to lava floated overa forest destroying flora and fauna. This after undergoing manygeological changes becomes carbonaceous material called Shilajit. It isabundantly available in Russia. It is the most widely used as medicinein the world. It is observed that although the general pattern of themolecules is found to be similar in Shilajit samples of differentsources, the variation in the conjugative properties of the molecules isfound different. This makes these medicines vary in their therapeuticefficacy hence, this type of fingerprinting is useful.

In the modern chemical analysis methods, determination of the percentageof active principles viz., alkaloids, flavonoids, enzymes, vitamins,essential oils, fats, carbohydrates, proteins, ash, acid-insoluble ash,and crude fiber is done by various analytical instruments. Some examplesshown below explain how standardization is done in modern science.

It is reported (WWW// Shilajit, Fulvic acid etc,. html) that, one of thevery important medicines used in Indian System of medicine Shilajit, isreported to have many compounds along with fulvic acids, and is claimedto be active principle. As this medicine from bituminous source iscollected from earth stored for many years, it can be seen that the moreit is in stored in the earth, the more it will be therapeuticallyactive. Nevertheless, in the entire globe, the geological variations maynot produce same molecules in all samples collected from different partsof the world. Another factor that influences the chemistry of thesedrugs is the purification process, which also needs to be standardized.

It is reported (WWW//Herbology. html) that, mostly standardization isdone for the individual key components, which have been empirically, andscientifically proven to be most advantageous for the human system. Sousually, standardization is done for certain molecules out of allpresent, which are found to have activity. However, the synergisticeffect of other compounds present in the medicine making the totalprofile of the medicine should be taken into consideration for itsefficacy.

It is reported (WWW//Tribulus Terrestrius puncture vine. html) that, thealcoholic extract of the fruits of Tribulus Terrestrius showsantiurolithiatic activity. In addition to this, the extract also shows asignificant diuretic activity. The alkaloid Harman has been reportedfrom the herb and Harmine from the seeds. The plant contains saponins,which on hydrolysis yield steriodal sapogenins. Many molecules offlavonoid in nature are reported in which the active principle is foundsaponins. The analytical report is given for the heavy metal analysisand total saponins content (20% w/w)

It is reported (WWW// Charak_com.—Quality control page—html) that, Humanlife is a synergy of Mental, Physical and Spiritual components, whichare related to the Indian Philosophy of Ayurveda where Pitta, Kapha andVata are considered as the basis for the total health of human beings.More details of standardization are given in the description oftraditional methods of therapeutic standardization.

It is reported (WWW//Standardized Herbal Extracts\ A herbalistsPerspective's, Dr. Micheal Tierra L. Ac.O.M.D html) that, due toEuropean Guaranteed Potency Law, it becomes compulsory to standardizeherbal medicines. The meaning of herbal standardization is defined asquantification of an active constituent or marker extract, where in theactivity is attributed to be the most.

It is explained (Frank R Stermitz et al, PAINS/February 15,200/Vol 97,No 4, pp 1433-1437) that, in the plant Berberis Aristata theantimicrobial property of the extract is due to the presence of5-Hydroxy Hydnocarpin, the berberin acting as anti microbial, andwithout which it will not. Hence, the synergistic effect of the entireconstituents should be taken into consideration while dealing with anherbal medicine but not only an active constituent.

WHO in its Regional Publication (Dr Ranjit Roy Choudary, Hrebal Medicinefor Human Health, Searo no 20) clearly mentioned what standardization isand what the member countries should do for the better use of itsmedicines by the people.

The role of acidity and alkalinity can be understood by carefullyunderstanding the extraction process of constituents from the medicinesat different pH values of the extraction solvents. This helps tounderstand the drug release mechanism in the intestine from the medicineconsumed by the persons having different intestinal pH. The role ofacidity and alkalinity was studied and understood carefully inunderstanding the therapeutic efficacy of a medicines. Acidity andalkalinity of organic and inorganic molecules are studied thoroughly toknow their properties as shown in Table 11 which appear later part ofthe specification. Of acidity and alkalinity shows the role of acidityand alkalinity on health (Health in Hands by Devendra Vora, Navaneetpublications (India) Ltd).

It is reported (WWW// Chewing. Html) that, in a study it was observedthat people with acidic systems absorb more pollution than people whohad established proper blood alkalinity. Acid/alkaline (pH) balance isimportant for normal cell function. More details were given in thearticle. Hence, the study of the acidity or alkalinity (organic orinorganic) in terms of ‘polarity’ will give the information of thetherapeutic efficacy of the medicines. Hence, the present method, whichcan do this job, will be of much use to know the therapeutic efficacy ofthe medicines. Using this proposed method, the acidity and alkalinitycan be established for the therapeutic standardization of medicines.

References made above will explain the conventional and reported methodsof standardization, wherein the individual constituents are isolated ata preparative scale and compared qualitatively and quantitatively withthe same compound present in the sample medicine under study.

In one of the reports (Pharmaceutical grade Saw Palmetto, Khwaja, et alU.S. Pat. No. 6,039,950) it was reported that the different individualfractions of ethanolic extract of Saw Palmetto were studied for theirbioactivity by measuring the determination of IC 50 in an androgenreceptor binding inhibition assay. Measurement of total fatty acid assayof whole extract and individual fractions were discussed. The fractionscontaining linolic acid ethyl ester and lauric acid ethyl ester wereidentified. The activity was calculated for each of the fraction forwhich androgen receptor binding inhibition has been assayed compared tothe total bioactivitty of the sample. The molecular weight and amount ofthe individual fatty acids were identified and incorporated in to thecalculation of bioactivity. The total bioactivity of the extract wascalculated comparing to the total percent activity of linolic and lauricacid ethyl ester fractions.

In the traditional medicine standardization, the total profile should betaken into consideration for the therapeutic efficacy of the herbalmedicine. Hence, in the present computer-based instrumental method, thetotal properties of all the constituents are taken into consideration assuggested in traditional concepts world over. The fingerprints of themedicines were proposed as a visual tool and proof for many purposes ofdealing with medicines particularly traditional. Before discussing themethod of the invention, the existing method of analytical method isgiven below.

Existing Analytical Methods of Chemical Standardization:

Improvement and use of modem analytical methods and instrumentation havedefinitely led to excellence in quality control methods of medicines.Improvement in analysis has led to more precise harvesting of many herbsas explained above and helped to prepare standardized extracts.

Although there are traditional methods of identification of medicinalplants like organoleptic, microscopic and physical, none of them givesan authentic identification, as given by a fingerprint of the plantmaterial, as far as the chemical profile is concerned.

Hence, it is proposed that the Chromatographic Fingerprint is muchuseful for quality control of medicinal plants instead of otherorganoleptic and microscopic studies. Since, ultimately it is thechemical constituents that are largely going to participate in thetherapeutic efficacy of the medicine, along with other properties of theherbal medicines; the analytical data of the chemical constituentsshould be able to provide the authentic efficacy of the medicine. It islike fingerprint of an individual gives the identity of him.

Till now Thin Layer Chromatography (TLC), High Performance Thin LayerChromatography (HPTLC) and High Pressure (Performance) LiquidChromatography are the methods commonly used for the analysis of anyorganic or organometallic compounds and finger printing. But, all themethods have some merits and demerits for an authentic analysis ofmedicines. The enclosed Table 12 appearing later part of the descriptioncompare various commonly existing methods of analysis, provide a generalidea of the merits and demerits of them.

FIG. 4, which is accompanying the specification, shows how thechromatogram developed from a TLC method is used as a “Chromatographicfingerprint” on the label of a commercial product. Except the assay ofconstituents no more information was given in the finger print.

After observing the above Table, it is found that the most suitabletechnique available for the analysis of a mixture of compounds is“Chromatography”, which gives the profile of the mixture after theseparation and identification with a suitable detector.

Out of the different types of chromatographic techniques available, thebest suitable is ‘High pressure liquid chromatography’ (HPLC). Althoughthin layer chromatography was used till recent times, advancementsbrought out in the equipment and separation columns of HPLC hasrevolutionized the analytical field of chromatography.

Most of the pharmaceutical analysis was reported in the form of achromatogram with the peaks due to molecules eluted by a mobile phasemostly reported in the official methods and pharmacopoeias. Theconstituents are analyzed after eluting on a HPLC separation columndetected by using any suitable detectors for analysis.

Usually the chromatographic analysis is done using a reference standard(internal or external). Without a standard reference material, theanalysis has no meaning because the peak of the chromatogram does notprovide any kind of chemical properties of the compound eluted. Hence,the confirmation of the qualitative and quantitative properties(spectral or chemical) of the components are unclear.

In the qualitative and quantitative analysis of medicines/drugs (Singleor Formulation), the emphasis is given mainly on the spectral andchemical properties of the components eluted after analyzing the sample.The analysis is done based on the influence of Electro magneticradiation on the analytes (say the UV-Visible radiation) and theirresponse to it. In the existing method of chromatography, the analyticalreport i.e., the chromatogram is not giving any of the chemicalproperties like polarity and UV-Visible absorptive properties of theconstituents. The chromatogram is not able to show the presence of themolecules which does not absorb at that wave length or have a different“Absorbance maxima” other than the set wavelength (say 225 or 254 nm).If the sample is 100% pure and if it is a known molecule then theanalysis at a fixed wavelength is acceptable, but it is highlyimpractical in the case of medicines where in more than one activemolecule are present. Some examples shown at single wavelength are givenin FIGS. 5-12, where in the chromatograms at various wavelengths aregiven. None of the single chromatogram is able to provide completeinformation about the chemical properties of the constituents present inthe medicine particularly in traditional medicines where more than oneactive principle may be existing. When the chromatograms and thefingerprints are compared, the utility of the fingerprints can be understood.

Hence, any chromatogram presented at a specific wavelength is not ableto provide the complete chemical profile of the ingredients present in asingle medicine and a formulation. So, the chromatogram is partial inits report, and is not acceptable. Any analytical method, which is notgiving complete information of the analysis, is not scientificallyacceptable.

In the analysis of herbal medicines, where different types of moleculesare present having different spectral properties (The absorbance maxima)the chromatogram at a fixed single wavelength will not be a meaningfulanalytical report or the chromatogram.

In the use of herbal medicines, the medicine as a whole is used withsome standard therapeutic conditions prescribed in the ancientliterature and scripts. Hence, the concept of searching for an activeingredient is said to be incomplete, because it is the total profilethat is responsible for the medicinal property of the medicine. So, anyanalytical method, which does not speak about the complete chemicalproperties of all of the constituents present in the medicine understudy, will not be useful.

Also the qualitative and quantitative profile of an herbal medicine varydue to many ecological factors like time of collection, place ofcollection, age of collection and monsoon conditions at the part ofcollection and soon.

It is already mentioned (Frank R Stermirtz et al,) that the synergy ofthe other constituents present along with the major constituent isequally important because the first will not be able to do its functionwithout the other constituents present in the extract as explained inthe beginning.

B. Prior Art on Therapeutic Standardization:

I) Traditional Method:

The great Indian Medical sages have understood and defined the conceptof Indian medicine by clearly defining the properties, constituents andhumors of the living beings. They also understood the inter and intrarelations amongst them. In almost all the traditional philosophies, thebasic concepts include the nature and its role on the humors of thehuman beings. It is said that the human body is made of seven types ofconstituents (Saptadhatus). The normal disorders (Tridosha) are of threetypes. The materialistic properties of any material in the universe aredue to five elements (Pancha bhutas). The interactions of differentpermutation and combination of these elements will influence the health.Hence, the understanding of these properties will help to understandtheir physical and chemical properties and so, there by theirtherapeutic efficacy. The philosophers in different parts of world havealso developed such concepts suitable for their tradition and society.

In ancient times (pre Samhitic and pre Susrutic period in India), thephysicians used Nadisastra (Science of reading pulse) to know the statusof the Tridosha (Vata, Kapha and Pitta) at the time of diagnosis to knowthe health status of the patient. The specific type of pulse (not theheart pulse) is studied to explain the type of disorder pre-dominant inthe patient (History of Medicine in India by Dr Priya Vrat Sharma,Indian National Science Academy).

It is used to understand the type of dosha(s) predominant in the patientat the time of diagnosis and the respective dosha(s) to be vitiated tocure the disorder. But this art of reading Nadi (Pulse) was confined tosome people of high caliber, personal skill and ability with lot ofdiscipline. Hence, every traditional practitioner was not able topractice it.

To over come this, the art of understanding the physico-chemicalproperties of the medicines and the humors of the human being had beendeveloped and standardized. The inter and intra relations of theseproperties with nature which influences health had been studied andstandardized thus the art of pharmacology and pharmaco-therapeutics wasdeveloped by the physicians.

The therapeutic efficacy of a drug is defined with use of a substancethat is capable of bringing about an (pharmacological) action in thehuman body (Kriyagunaval) and due to the collective functioning of manyfactors, (samavayikaranam), just as a piece of cloth results becausefrom its many component threads acting together.

In the world, there are two main types of living things, the plants andanimals. It is also said that this world is made of five great elementsi.e. Earth, Water, Air, Fire and Space (as said Panchabhutas inAyurveda). The basic properties of these materials are of two types,namely Strong (Powerful) and Mild (Soft). If we accede to this highlytenable logic, we can say that in this world, all actions are due todifferent permutation and combination series of the above properties,giving a wide range of properties and materials varying in theirintensity.

In the philosophy of most of the traditional medicine world over, theco-inherence of the nature of the five constituents is taken intoconsideration by which the body is made. They will help in understandingthe disease or disorder of the patient. This coherence is called Purushain Ayurveda, Yin and Yang in Chinese medicine. Table 6 appearing laterpart of the description gives how the Chinese system has used the abovetwo factors, how they were classified and defined to standardize thetherapy and diseases.

Chinese medicine classifies the status of the human body as Yin and Yangrepresenting sorrow and happiness as mentioned above. These factors areattributed for various properties of the medicines and living beings.The maintenance of these factors is done holistically by taking the roleof chemical, physiological and social factors in to consideration. Mostof the time the Chinese medicine has a direct or indirect relation withvarious bio energy centers located in the body. The art of acupunctureuses the same. The other factors that reported in other philosophieshave resemblance with Chinese medicine. After the panchabhautic concept,the concept of Tridosha (Pitta, Kapha and Vata) plays a major role inthe Indian traditional medicine and the seven constituents (Saptadhatus)by which the body is made up of Ayurveda believes in the holisticphilosophy of life and emphasis is given for the prevention of diseasesrather than curing of diseases.

The holistic approach of ayurveda advocates that the soul, mind and thebody are the three integral parts of life and when these are in dynamicequilibrium and harmony, the state is called good health (Arogya). Whenthey are in inequilibrium and disharmony, the state is called disease.(Vaishamya).

According to Ayurveda, Tridosha maintains the physiological features ofvarious systems in dynamic equilibrium status. In other words, harmonyof tridoshas bestows good health, disharmony results to disease. Hence,most of the time the tridoshas are dealt with, in curing any disease.The selection of drug is made for the disease that should be dealt with.

A disease is defined as “Any thing that brings a sadness and grief tothis person (Purusha) is a disease. They are of four types 1.Theaccidental (agantavaha) 2.The body born (Sarirah) 3.The Mind born(Manasah) and 4. The natural (Swabhavikah). It is for this reason, mostof the traditional concepts deal with both psychosomatic factors to curethe disease along with a disciplined and standardized method of life.

The diseases were classified into three classes generally. 1. Thecurable (Sadhya) 2. The Mitigateable or manageable (Yapya) and 3. Theincurable (Asadhya). As said above, it is mostly considered as thosebodily diseases having their source arise by the incompatibilities ofthe thridoshas viz., Vata, Kapha and Pitta and blood individually or incombination with one another. But, the diseases that arise not due tothe above reason like psychological are dealt in a different way. Thatis why any traditional concept is used to take all the psychosomaticfactors in to consideration to deal with a disease. The individualproperties of the doshas are explained as given below.

Broadly it is outlined that the Vata or vayu dosha deals withendocrinological, neuromuscular and nervous activities all those thatcause the major or the gross dynamics of the life, the foods that causegas formation can be classified in this category. Pitta dosha refers todigestion and chemical functions or rasa kriya in general and Kaphadosha includes factors providing form, stability, and cohesion andlubrication factors. As the first dosha, ‘Vata’ is considered toinfluence the other two it is considered as the key factor for anydisease. An elaborate description of these humors is given in elsewherein the body of the text.

The decrease of vata leads to general dullness in activity. Hence, thedrugs, which decrease this dullness, will be of Vata Hara. The decreaseof digestive capacity is called as pitta dosha. A medicine, whichincrease the digestive capacity or activate bile mechanism, will be ofPitta Hara in nature. The decrease of liquidity or mucous will lead toroughness, an internal burning, an emptiness in stomach, a looseness inthe joints, thirst, weakness and a continues insomnia. These are thebasic symptoms of kapha disorder. Any medicine that vitiates thisdisorder will be of Kapha Hara in nature.

A detailed description of all the factors is given for variousphilosophies in order to under stand more generally about differenttraditional medicines world over. Table No 1 and Table No 2 (which areappearing in the later part of the description) gives an elaboratedescription of the Indian Ayurvedic philosophy and various components init.

Hence, to understand the therapeutic efficacy of a medicine or food, oneneeds to understand their physical and chemical properties. In theancient times people use to understand these properties using theorganoleptic methods like the taste, the smell and the color of thematerials. The basic properties classified were 1. Taste (Rasa), 2.Quality (Guna) 3. Potency (Virya) 4. Post assimilative status and effectof the constituents (Vipaka) and 5. Special action (prabhava,geometrical and optical isomer molecules)

It is these three factors namely, the Doshas (Disorders), the Dhatus(constituents) and the Malas (excreta) that are mainly to be dealt forcuring a disease or a disorder. If the above-mentioned properties of themedicines tally with the dosha, it will be vitiated or neutralized, thusthe disease is cured.

The classification and differentiation of drugs according to Ayurvedicpharmacodynamic and genetic principles vary from one situation toanother according to doshic predominance of the patient. In other wordsthere is a relation between the dravya gunas (medicinal properties) anddoshas (disorders). Addition or deletion of one or more drugs may benecessitated to treat an identical disease with the patients withdifferent individual doshas or combination of doshas. Hence, Ayurvedicpharmacotherapy is more individualistic according to dosha predominanceof the patient and not generalized as in the case of modern medicalpharmacotherapy. Identification of properties (Rasa, Guna, Veerya,Vipaka and Prabhava) compatible to doshas is unique and more reliable inAyurvedic pharmacotherapy.

ii) Modern Method of Therapeutic Standardization:

The existing pharmacotherapy has not taken the above mentioned conceptsinto consideration. Phytochemists are interested only in isolation,purification and structural elucidation of the active principlesisolated from the plants and they passed on them to pharmacologists tostudy their biological activity. The pharmacologists in turn screen themolecule(s) for pharmacological activity, establish its mechanism(s) ofaction and substantially rate its efficacy in comparison with theexisting standard drugs used in modern medicine.

This concept is in no way going to help the traditional medicalpractitioners since the isolation of the active principle(s) drasticallychange the holistic character of the medicines and their therapeuticefficacy.

Instead of assaying the solvent extraction fractions, active principlesetc., obtained from the individual plants, the analysis of total extractfrom a medicine using a solvent compatible to the human cells and cellmembranes of the body will be of much use to evaluate thepharmacological activity of such medicines.

In the modern clinical trials conducted for the therapeuticstandardization, they are done in three phases (four in the case ofinternational utility), involving large number of people. Theinformation regarding a new medicine to be submitted to Drug Controllergenerally consists of:

-   1. Chemical structure-   2. Pharmacological class-   3. Formulation details-   4. Data on animals including data on toxicity studies-   5. Data on clinical pharmacology including pharmacokinetics    (Behavior of the drug in the human body)-   6. Pharmacodynamics (Actions of the drug inside the body)-   7. Special studies and status of the drug in the rest of the world.-   8. Data on Bio-Equivalence studies

The Phase one study is mainly concerned with assessing the drug's safetyto know how the medicine is absorbed, metabolized in the human body andexcreted, also it envisages to estimate the side effects and the dosage.

The phase two studies are dedicated to test the efficacy in a randomizedway. One group of patients will be given the actual medicine and thesecond with placebo.

In the phase three study, a large scale testing will be taken up tostudy the effectiveness, benefits and the range of possible adversereactions of the drug. After successful completion of this step, theindustry will go for marketing the drug.

In late phase III and IV studies, pharmaceutical companies will haveseveral objectives. Studies will help to know the efficacy of the newdrug compared to an existing drug. The long-term effectiveness andimpact on a patient's quality of life due to the new drug will be known.The cost effectiveness of the drug therapy relative to other traditionaland new therapies will be known.

But all the above studies are costly and time consuming. They will notbe taking into account of the role of the ecological factors, thegenetical discipline (as practiced in the Indian family and marriagerelations), the psychological, the social and other variable parametersof the patient in to consideration. This will make the effectiveness ofthe drug limited to a particular group or genetic type of people.

C. Prior Art of Barcoding and Enterprise Resource Planning (ERP)Customer Relationship Management Applications:

The modern method of making any commercial goods proprietary isBar-coding. For all commercial transactions, the barcode is widely usedin many ways. To make the medicines identified as proprietary goods, anovel method of bar coding is proposed in this invention.

It is reported (Peernet bar-code store (Java Active X servlete-business)) that 1800 character and 2700 digits (Even 9,99,999 numbers)can be fed to a commercially available bar coding software to generate abarcode of any item. When a digital value and/or numerical number isgiven to the bar-coding software it generates a specific barcode patternby the logic specified in the software proprietary for a user.

The barcode thus generated will present and display the attached“display window” file information having all details of theproduct/label, when a barcode readable vending machine sees (throughelectronic eye or sensor) the barcode on the product. The barcode can beread from any ERP and CRM applications world wide through network.Presently the catalogue numbers are being used for bar coding themedicines and related products which does not specifically contain anychemical property of the product as proposed in the method.

Objectives of the Present Invention

The main objective of the invention is to propose a novel method ofchromatographic finger printing, chemical and therapeuticstandardization and bar coding of organic and organo-metallic moleculesfrom a plant, animal or a naturally available or man made materials usedas medicines.

Another objective of the present invention is to provide a novelchromatographic finger printing of herbal medicines and formulationswhich obviates the drawbacks detailed above.

Another objective of the present invention is to provide a completechemical analysis of the constituents present in the medicine understudy and their conjugative properties indicating the therapeuticefficacy as per the traditional concepts of the medicine using newsoftware developed.

Still another objective of the present invention is to provide a novelmethod for chromatographic fingerprinting of herbal medicines useful forthe quick identification of the actual profile of the compounds presentin the medicine under use along with their therapeutic efficacy of theconstituents.

In yet another objective of the present method is to provide a novelchromatographic finger printing of herbal medicines and formulationsusing the contour and 3-D chromatograms of the herbal medicines andformulations.

In yet another objective of the present invention is to provide a novelmethod for chromatographic finger printing of herbal medicines useful tocheck the adulteration of the compounds present in the medicine underuse.

Yet another objective of the present investigation is to prepare astandard analytical parameters like extraction with same solvent ethylalcohol, same run time 0-60 min, same mobile phase acetonitrile alongwith phosphate buffer having a pH in the range of 5.5-7.5, and a sameUV-Visible Range of 200-800 nm.

Yet another objective of the present invention is, to categorize andquantify the constituents of a medicine based on polarity andconjugation from 3-D and contour chromatograms and assess thetherapeutic efficacy of the medicine on which humors it is going to act(vitiate).

Yet, another objective of the present invention is to provide a barcodefor the selected peak of a molecule given in the image.

Yet another objective of the present invention is to prepare a databaseof barcodes for the fingerprints developed useful for all types ofdatabase applications.

Yet another objective of the present investigation is to generatedisplay windows for all the samples of the fingerprints having thedetails of the samples like 3-D and Contour fingerprints, the barcode,details of the origin (Industry or Country), manufacturing date, date ofexpiry, reported dosha, individual constituents used, their assay, batchnumber, lot number, M.R.P (maximum retail price) etc.

Yet, another objective of the present invention is to attach the displaywindows with the respective barcodes, facilitating to deal with displaywindows in all applications whenever they are used as a source of dataand information.

Yet another objective of the present invention is, to prepare a databaseof display windows thus generated and attached to the respectivebarcodes, to use in the Enterprise Resource Planning (ERP) and CustomerRelationship Management (CRM) applications for all commercial networkingtransactions of the medicines and samples.

Yet another objective of the present investigation is to prepare adatabase of barcode and display windows and any information, speciallyrequired for the regulatory authorities to control the movement of themedicines in and out of the country.

Yet, another objective of the present invention the UV-Visible spectraof the compounds will provide the conjugative properties of themolecules and the concentration of the individual concentrations of themolecules along with the polarity of the molecules.

Yet another objective of the present invention the use of finger printof contour and 3-D chromatograms will be the basis for theidentification of chemical constituents to limit the scope of theinvention.

Yet another objective of the present invention is to develop a method offingerprinting for the adulterated food and drug samples, substitutedand contradictual food and drug samples and commercial samples of foodand drug samples and to identify the pure and adulterated.

Yet, another objective of the present invention is to develop a methodof fingerprinting for the organic and organo-metallic constituents inany type samples to identify the chemical constituents present in it forvarious purposes of quality control and process standardization.

Yet another objective of the present invention is to develop a method offingerprinting for the Allopathic, Ayurvedic, Homoeo, Siddha, Unani,Chinese, Tibetan and Kampo (Japanese) medicine samples for the qualitycontrol and chemical and therapeutic standardization

Yet, another objective of the present invention is to develop a methodof fingerprinting for the study of variation of chemical constituents innaturally occurring or synthetically prepared samples and to identifyand standardize the chemical constituents in them.

Yet another objective of the present invention is to develop a method offingerprinting for the study of variation of chemical constituents innaturally occurring or synthetically prepared samples and to identifyand standardize the variation in chemical constituents in them due toGeological, Ecological, Genotypic and Phenotypic variation factors.

Yet, another objective of the present invention is to develop a methodof fingerprinting for the study of chemical constituents in herbalproducts of single and formulated medicine samples and to identify thechemical constituents in them for chemical and therapeuticstandardization.

Yet, another objective of the present invention is to develop a methodof fingerprinting for the study of variation of chemical constituents inbiological samples and to identify and standardize the chemicalconstituents in them.

Yet, another objective of the present invention is to prepare a largedatabase, which will give many generalizations of the therapeuticefficacy of a particular group of plants, classified as a group for aparticular disease or therapeutic classification.

Yet another objective of the present invention is to provide a methodwhich enables to understand and standardize the Physico-Chemicalproperties of the medicines like color used in traditional method oftherapeutic standardization using conjugative and polarity properties ofthe individual constituents and the whole medicine.

Yet another objective of the present invention is to provide a methodwhich enables to understand and standardize the Physico-Chemicalproperties of the medicines like Taste (Rasa) namely Sour, Salty,Pungent, Bitter, and Astringent (Amla, Lavana, Katu, Tikta, and Kashayaas said in Ayurveda respectively) used in traditional method oftherapeutic standardization using conjugative and polarity properties ofthe individual constituents and the whole medicine.

Yet, another objective of the present invention is to provide a method,which enables to understand and standardize the Physico-Chemicalproperties of the medicines like Quality, Potency, Metabolite afterassimilation or such modifications and Specific properties likeChirality of the molecules (Guna, Veerya Vipaka, and Prabhavarespectively as said in Ayurveda)

Yet another objective of the present invention is to provide a methodwhich enables to understand and standardize the Physico-Chemicalproperties of the medicines like Heavy, Light, Cold, Hot, SoftLubricated Supple, Dry, Slow, Sharp (Guru, Laghu, Sheeta, Ushna,Snigdha, Manda, Teekshna respectively as said in Ayurveda) used intraditional method of therapeutic standardization using conjugative andpolarity properties of the individual constituents and the wholemedicine.

SUMMARY OF THE INVENTION

This invention relates to a method for detection and identification ofprinciples from extracts of plants or animal, natural or syntheticsources, using chromatographic finger printing techniques, said methodcomprising the steps of:

-   i.) extracting the organic or organo-metallic molecules using a    suitable solvent;-   ii.) subjecting the extract obtained in step (i) to the separation    analysis, using High Pressure Liquid Chromatography techniques;-   iii.) generating contour and 3D chromatograms of the ingredients    eluted based on the pH and polarity;-   iv.) converting the 3-D and contour chromatogram obtained into a    colored image, analyzing the colored image for its individual colors    using the co-ordinates denoting all its 3-dimensional properties of    the said image by using an in-built software;-   v.) denoting the concentrations of the various constituents eluted    with time;-   vi.) generating a chromatogram based on color analyzed, having peaks    at various retention times along with conjugative properties of the    molecules;-   vii.) identifying the compounds in the said ingredients by the    UV-Vis absorptive properties of the various constituents in the    image;-   viii.) identifying, determining and classifying the compounds eluted    as polar, medium polar and less or non-polar based on the polarity    and conjugative properties;-   ix.) generating a barcode for a selected peak using the X axis as    Retention Time, Y axis as Wavelength, R as number of Red Pixels, G    as number of Green Pixels and B as number of Blue Pixels; and-   x.) generating a database of fingerprints and barcodes and    identifying the respective compounds in the samples.

DETAILED DESCRIPTION OF THE PRESENT INVENTION

Accordingly, the novel basis of the present method is, presenting thespectral properties of the chemical constituents displayed in 3-D andcontour chromatograms as a novel method of fingerprinting. Thechromatogram generated by this method provides the conjugative andpolarity properties of the individual molecules present in the medicinesgiving the therapeutic efficacy of the medicine.

In a molecule the UV-Visible absorbance capacity of the moleculesdepends on the structure of the molecules. When the double bonds ortriple bonds are present in the molecules alternatively in thestructure, it is called as conjugated. The more the molecule isconjugated the more it will be chemically and biologically active. Hencethe more the molecule is conjugated the more it will be therapeuticallyactive. Thus, the measurement of conjugative properties will give thetherapeutic efficacy of a medicine. Hence, the use of conjugate propertyfor the therapeutic standardization is the novelty of the presentinvention.

A novel method is proposed for the quality control of herbal medicinesand formulations mostly useful for the fingerprinting andstandardization (chemical and therapeutic) of Traditional Medicinesunlike a method being used for analyzing only active ingredient (whichis not known in many herbal medicines) for the analysis of medicines ata single wavelength. It gives the total profile of the chemicalconstituents present in the traditional medicines along with physicaland chemical properties of the compounds (Say UV-Visible absorptiveproperty and polarity property). In the first part of the method, animage of the fingerprint of the medicine will be generated. But as imagecannot becomes analytical data, a computer-based method is developed togive the qualitative and quantitative data of the ingredients in theform of an analytical chromatographic report. The same is proposed as anovelty of the method of the invention.

The reactivity of any molecule will depend upon the number of double andtriple bonds existing in the molecules along with the electrophilic andnucleophilic sites on the molecule. The moieties donating electron andaccepting electron will create difference in the total electrical chargeof the molecule. This makes the molecule polar. Hence polarity of themolecules will provide information about the capability of a molecule todonate or accept the electron with another molecule. This will controlthe activity of a molecule. Thus, the information of the polarity of amolecule will speak about the reactivity of the molecule. In the presentmethod, the chromatogram provided by the method will give theconjugative and polarity properties of the constituents present in amedicine in the finger print. Thus, this method is used for thestandardization of the medicines to know the therapeutic efficacy of amedicine using their conjugative and polarity properties of themedicines. This is the novelty of the proposed method.

As said above the UV-Visible spectra and polarity of the compounds willindicate the conjugative and polarity properties of the compounds andthus indicating the chemical/medicinal property of the medicines. Thisprofile of spectra of all the constituents in a single picture, “thefinger print” as proposed now will become the blue print of theconstituents present in herbal medicines and formulations. This becomesa superior method of identification and standardization of herbalmedicines than the existing, as the peaks will express the UV-VIS.Properties or conjugative and polarity properties of the constituents,unlike in a conventional chromatogram taken at a single wavelength alongwith the quantification of the constituents.

As described in the traditional standardization methods the colors ofthe medicines were used to know and standardize their therapeuticefficacy. The colors of the molecules are understood by their absorptiveproperties of the radiation of the UV-Visible range of radiation. Basedon the structure, functional groups, conjugation, and the extent ofunsaturation the absorbance of a particular wavelength depends. The morethe molecule is conjugated the longer the wavelength of absorption willbe. Hence, the UV-Visible absorbance of any molecule is widely used inthe qualitative and quantitative properties of the constituents. Thecolors and the therapeutic efficacy of various medicines were given inthe ancient literature.

Ultimately the colors of the molecules are due to a specific chemicalnature of the molecule. When the same is studied, the chemical propertycan also be understood. In ancient times the colors of the flames wereused for the quality control of metals and related products this,involves the basic spectrophotometric principles. Thus, study andunderstanding of the interaction of the electromagnetic radiation willbe useful to study the chemical nature and thus the therapeutic efficacyof the medicines. The same principle has been used in the presentspectrophotometric method of fingerprinting and standardization. Themain novelty of the present method involves in the “division of thefingerprint into different therapeutic zones based on the scales ofwavelength (Conjugation) and retention time (Polarity) to understand thetherapeutic efficacy (in traditional terms) of a single or a formulatedmedicine” using an instrumental and software based program.

Using the computer-based software developed, a barcode is generated fora selected peak of a molecule given in the image. Where, X is theRetention Time, Y is the Wave length in contour chromatograms andabsorbance in 3-D chromatograms, R the red color indicating the highestconcentration of the constituent, G the green color indicating thelesser concentration of the constituent and B Blue color indicatingstill lesser concentration of the constituent are the coordinatesprovided by the present software, is feed in any commercially availablere-salable bar coding software, added in the present software generatesa barcode for a single constituent, or for many constituents. The Imageof the fingerprint is viewed on a display window attached to it. Thiswill be displayed whenever the electronic eye of the vending machinereads the barcode. This makes the image (Finger print) and barcodeproprietary for a product of an industry or a country. This is claimedas another novelty of the method of the invention.

When the polarity of the column is fixed and the polarity of mobilephase is varied constantly in an increased or decreased order, on areverse phase column, the constituents present in the sample will elutein the order of high polar constituents eluting first the medium polarconstituents next followed by non-polar constituents. Care is taken ineluting the constituents in the order of increased or decreased order ofpolarity such that no constituent of any polarity will be left un-elutedfrom the column achieving total elution. The order and properties ofpolarity and elution in the case of normal phase columns are applicablesame as in the case of reverse phase column but in reverse. In a normalphase column the non-polar constituents will elute first and followed bypolar constituents, based on order of polarity of the mobile phase usedfor elution.

Thus, a fingerprint developed having the chemical constituents arrangedin the increased or decreased order of polarity will help to bringtherapeutic generalizations about the medicines. This is another noveltyof the proposed method.

The image of contour chromatogram developed after the analysis isdivided in to three zones on X and Y-axis. The conjugative property(Absorption of a particular wavelength of radiation) is taken on Y-axisand polarity is taken on the X-axis, as the elution of the constituentsis controlled using the polarity of the mobile phase composition. Now asreported in literature the Y axis is scaled as per, the therapeuticefficacy based on wavelength (color). The entire image is divided in tosix chambers wherein the chemical constituents have specific conjugativeand polarity properties. This in turn is proportional to the therapeuticefficacy of the constituents in the chamber. Thus when a medicine isfingerprinted, based on the color represented for the absorption of aspecific wavelength and having a specific polarity, the total colors inthat zone is calculated and interpreted for the therapeutic efficacy ofthe constituents present in it. Thus, the holistic therapeuticstandardization and chemical standardization is achieved using thismethod.

Mostly the elution of the samples were done from high polarity mobilephase to low polarity mobile phase. Thus in the finger prints theconstituents present in the first zone (Zone-1) will be of high polar innature. The same pattern applies to the other zones, the medium polarconstituents eluted in the medium polar zone (Zone-2) and the low ornon-polar constituents eluted in the non-polar zone (Zone-3). Thispattern reverse when a normal phase column is used due to its elutionproperty as described above.

Most of the high polar molecules are highly reactive chemically, thusbiologically. When they enter the first part of the digestive systemmouth, they will immediately start acting on the biological system andthe enzymes present there. Then the constituents will enter the stomachand intestine where they will under go different changes (Postassimilation effects, Vipaka in Ayurveda) due to the digestive juicesand their enzymes present in the part. In the process of absorption, themoment the molecules of high activity (high polar) immediately startinteracting with the biological system and show their therapeuticproperties. This is compared that in Ayurveda, the intestinal part ofthe human body is classified as Pitta zone, where the high polarmolecules are playing a major role. The heat causing mechanism will playan important role in the diseases and biological mechanisms related to.It indirectly indicates the molecules of high reactive, the high polarmolecules. After the absorption, the blood with all the absorbedconstituents will carry them to heart and the parts related to it. Thenthe blood will be sent to different parts of the body. In Ayurveda, theupper portion of the human body is defined as the Kapha zone, where thecold mechanism will be playing an important role. Thus, the molecules ofmedium polar molecules will play an important role in the mechanismsrelated to this zone.

The low polar and non-polar constituents will be able to enter to thehuman body only through blood transfer, Thus the body organs where themechanism of availability of the chemical constituents is only by blood,will be coming in the last category of the polarity. The non-polar oils,fats and other such molecules and mechanisms in the human body areclassified as Vata disorders and all such disorders are cure using thesame type of materials.

The low and non-polar constituents will be eluting in the last zone ofthe fingerprint. Thus, this zone (zone-3) is considered as Vata zone.Thus, the basic humors of the molecules can be identified based on theirpolarity, which facilitates to know on what disorder (dosha) it is goingto act upon. Thus, the present method is useful for the therapeuticstandardization of the medicines.

Thus the total constituents present in the Zone-1 Pitta zone, Zone-2Kapha zone, Zone-3 Vata zone are present in the form of a pie diagramwhich represents the ratio of the efficacy of the medicine on each ofthe disorder. Thus, medicines having constituents in the order of50:20:30 will be medicines of tridoshahara of the order of 50%: 20%:30%. Thus, the therapeutic efficacy is standardized quantitatively. Theincrease or decrease of any one or two of the other doshas is done byformulating medicine by adding other medicines and prepare a suitableformulation needed to cure a specific individual.

Thus, a fingerprint having the scales of conjugation, absorbance andpolarity the 3-D chromatogram will give information about thetherapeutic efficacy of the medicine. Analyzing it using all its threedimensional properties of the said image will do quantification of 3-Dchromatograms of the medicine. For example if the 3-D chromatogram isconsidered as a ‘cap with a hood’ the matching of the entire cap 3dimensionally, with another sample of different qualitative andquantitative properties, the extent it matched will be presented as ananalytical report qualitatively and quantitatively. Here the hood of theCap is compared to the peak of the molecule at a particular wavelength.A sample with more number will like a cap with many hoods. Thus thematching of the three dimensional coordinates will provide a foolproofmethod of comparison and analysis. The coordinate it matched will givequalitative and the extent it matched will give the quantitative data ofthe sample understudy. This is made possible by special softwareprepared for this purpose. This becomes an ultimate method of qualitycontrol. This is another novelty of the method of the invention.

The invention also relates to a software based data processor of 3 Dchromatograms and color contour image of an ingredient, said processorcomprising computing means and capable of:

a. an analyzer (extracting colors) for analyzing the colored contourimage based on the selection of various colors (with standards mentionedin release notes, life cycle, processing) denoting the concentrations ofthe various constituents eluted with time, and polarity based onretention time;

b. an analyzer for analyzing the 3-D chromatograms of the medicine usingall its 3 dimensional properties of the image;

c. a means for generating a chromatogram having peaks at variousretention times along with conjugative properties of the moleculeseluted with time in a specified order of polarity;

d. an identifier for identifying the compounds in the said molecules bythe UV-Vis absorptive properties of the various constituents in theimage;

e. a means for correlating the reported biological, therapeutic activityof the of various constituents present in the medicines understudy basedon the polarity and the conjugative properties of the molecules bydividing the fingerprint into therapeutic zones on X and Y axis;

f. a means for generating a barcode for a selected peak(s) using theimage coordinates viz., X for retention time, Y for wavelength, R fornumber of red pixels, G for number of green pixels and B for number ofblue pixels, provided by the proposed software;

g. a means for generating a database of fingerprints and barcodes forthe samples, facilitating all kinds of database utilities likeEnterprise Resource Planning (ERP) and Customer Resource Management(CRM) applications; and

h. a means for generating a database of the ‘display widows’ for all thesamples to be used by the ENTERPRISE RESOURCE PLANNING (ERP) andCUSTOMER RELATIONSHIP MANAGEMENT (CRM) type of business applications.

Abbreviations Used in the Patent Document

-   1. ERP: Enterprise Resource Planning-   2. CRM: Customer Relationship Management-   3. UV-Visible: Electromagnetic radiation in the range of 200 nm to    800 nm-   4. Organic molecule: A molecule having basic elements of C, H, N, O,    S in its structure.-   5. Organo-Metallic molecule: A molecule having a metal along with    basic elements of C, H, N, O, S in its structure-   6. Contour Chromatogram: A type of chromatogram displayed in the    data generated from a Photo Diode Array detector, which scans the    sample with electromagnetic radiation in the range of 200 nm to 800    nm. The chromatogram thus generated will provide Retention time on    X-axis, range of absorbance (nm) on Y-axis. Different colors will be    used to represent different concentrations of the individual    constituents.-   7. 3-D chromatogram: This is also generated using the same set of    equipment as given above. It will be more informative providing the    UV-Vis spectra of each of the in-gradients after separation from a    mixture. It helps to identify the constituents using the spectrum.-   8. Ayurveda: An Indian philosophy written by Indian sages explaining    organized science of medicine and health discipline.-   9. Oshadisukta: a chapter in Rigveda giving the details of the    properties of the medicines used as medicines.-   10. Rasa, Guna, Veerya, Vipaka, and Prabhava: Different    Physico-Chemical properties of the medicines and materials used to    understand the efficacy of the medicines used in Indian system of    medicine.-   11. Lokapurusha Samanya: Law of Uniformity of Nature-   12. Tri doshas: three humors using which the human body is studied    viz., Pitta, Kapha and Vata used in Indian system of medicine.-   13. Prakriti-Purusha: The first one is compared to Mother Nature    (woman) and the second to person (man) used in Indian system of    medicine-   14. Pitta: A term used in Indian system of medicine for one of the    humors in the bddy in Indian system of medicine to denote the    personality or a disease which denotes the digestion and chemical    functions or rasa kriya in general in the human body.-   15. Kapha: A term used in Indian system of medicine for one of the    humors in the body in Indian system of medicine to denote the    personality or a disease which denotes the factors providing form,    stability, and cohesion and lubrication factors in the human body.-   16. Vata: A term used in Indian system of medicine for one of the    humors in the body in Indian system of medicine to denote the    personality or a disease which denotes the neurological and    endocrinological, and nervous activities in the human body.-   17. Geological factors: Global variation in soil nature and ground    water constitution etc related to earthly components.-   18. Ecological Factors: Global variation in tropical regions,    monsoon conditions and temperatures.-   19. Organoleptic methods: Methods of identification of the    properties of medicines like 1. Taste (like Sour (Amla), Salty    (Lavana), Pungent (Katu), Bitter (Tikta), Astringent (Kashaya)) 2.    Color, 3. Odor and 4. Texture etc., using human sensory organs.-   20. Tastes (Rasa Physico-Chemical properties) Properties visibly    seen (Color, Size) and felt (Texture) and all physical properties    and properties like taste and medicinal related to the chemistry of    the individual constituents present in the medicines.-   21. The medicines were standardized using their properties like    Taste (Rasa), Quality (Guna) Potency (Virya), Post assimilative    status and effect of the constituents (Vipaka) and Special action    (Prabhava,)-   22. Saptadhatus: The 7 elements like Rasa (Body fluids), Rakta    (Blood), Mamsa (Muscle), Majja (Bone marrow), Asthi (Skeleton    System), Medas (Fat) and Shukra (Reproductive) constituents present    in the human body used in Indian system of medicine.-   23. Panchbhutas: The 5 natural elements like Prithivi (Earth), Ap    (Water), Teja (Fire), Vayu (Air) and Akasha (Space) present in the    world used in Indian system of Medicine-   24. Nadisastra: A science, which explains the health status in the    human body, used in Indian system of medicine by studying the pulse    of the person.-   25. In Indian System of Medicine, Factors causing disease are    explained as Agantavaha (accidental), Sarirah (body born), Manasah    (Mind born) and Swabhavikah (natural)-   26. The diseases were classified into three classes as per Indian    System of Medicine They are Curable (Sadhya), Mitigateable or    manageable (Yapya), The incurable (Asadhya)-   27. Conjugative property: If a molecule has alternative single and    double bond and electron donating and accepting property it is    called conjugative. This is seen in the UV-Visible spectrum of a    molecule. Based on the energy absorbed for the excitation of Sigma    and Pi electrons in a molecule from the electromagnetic radiation,    the molecule will absorb a specific wavelength of radiation. The    absorbance maxima of a molecule will thus indicate the conjugative    property of the molecule under study.-   28. Polarity property: If a molecule has difference in its    electrochemical property, it is called polar. This depends on the    atoms attached to the molecule with electron donating (nucleophilic)    or electron accepting (electrophilic) moieties or functional groups    the molecule will have a difference in its electrical charge on its    molecular orbital. This makes the molecule to have a positive end    and negative ends. This type of molecules is called polar molecules.    The extent and type of electrical charge will make the molecule    polar, medium polar and non-polar in nature.-   29. Gradient or ternary system of HPLC: A HPLC instrument having two    or three liquid pumps to vary the ratio of the aqueous or    non-aqueous solvents. This will help to control the total polarity    of the mobile phase as per the requirement.    Some abbreviations used in software:-   1. JDK: Java Development Kit-   2. Con: Contour Chromatogram-   3. 3-D: 3-Dimensional Chromatogram-   4. WOS: Without Scale-   5. X: Represents the Retention Time of the chromatogram-   6. Y: Represents the absorbance in the 3-D chromatogram and wave    length range in contour chromatogram-   7. R: Intensity of red color at a particular pixel position-   8. G: Intensity of green color at a particular pixel position-   9. B: Intensity of blue color at a particular pixel position

EMBODIMENTS

One of the present embodiments of the present invention relates to amethod for chromatographic finger printing, chemical and therapeuticstandardization and bar coding of organic and organo-metallic moleculesfrom a plant, animal or a naturally available or man made materials.

Another embodiment of the present invention relates to a novel methodfor chromatographic finger printing of herbal medicines andformulations, which obviates the drawbacks detailed above.

Still another embodiment of the present invention relates to a methodfor a complete chemical analysis of the constituents present in themedicine under study and their conjugative properties indicating thetherapeutic efficacy as per the traditional concepts of the medicineusing new software developed.

Still another embodiment of the present invention, relates to a methodof novel chromatographic finger printing of medicines that is useful forthe quick identification of the actual profile of the compounds presentin the medicine under use along with their therapeutic efficacy of theconstituents.

In yet another embodiment of the invention, an embodiment of a novelconcept of chromatographic finger printing of herbal medicines andformulations using the contour and 3-D chromatograms of the herbalmedicines and formulations is proposed. They were developed on a PhotoDiode Array detector (PDA) of a High Pressure Liquid Chromatograph. Thisdelineates the data of the spectral properties of the constituentspresent in the herbal medicines eluted under experimental analyticalconditions.

In yet another embodiment of the present invention relates to a methodfor the chromatographic finger printing of extract from a medicine ofany nature containing molecules that absorb Ultraviolet and Visiblerange of radiation (200-800) or of any range of electromagneticradiation.

In another embodiment of the present invention, the UV-Visible spectraof the compounds provide the conjugative properties of the molecules andthe concentration of the individual concentrations of the molecules.

In another embodiment of the present invention the fingerprintsdeveloped for a same medicine extracted under different pH values arehelpful to understand the drug release in the intestine system atdifferent pH values of an individual.

In yet another embodiment of the present invention, the UV_VIS spectraof all the constituents are shown in a single image “The ChromatographicFingerprint”

In yet another embodiment of the present invention the finger printbecomes the blue print of the constituents present in a herbal medicineor formulation for an assay and quick identification of the medicineunderstudy.

In yet another embodiment of the present invention, the fingerprintusing the contour and 3-D chromatogram is the basis for theidentification of chemical constituents existing and/or formed new.

In yet another embodiment the UV-Vis spectra and polarity of thecompounds indicates the conjugative and polarity properties of thecompounds and thus indicating the chemical/medicinal property of themedicines. This profile of spectra of all the constituents in a singlepicture, “The Finger Print” as proposed now becomes the blue print ofthe constituents present in herbal medicines and formulations. Thisbecomes a superior method of identification and standardization ofherbal medicines than the existing, as the peaks express the UV-VIS.Properties or conjugative and polarity properties of the constituents,unlike in a conventional chromatogram taken at a single wavelength alongwith the quantification of the constituents.

In yet another embodiment of the present method the “division of thefingerprint into different therapeutic zones based on the scales ofwavelength (Conjugation) and retention time (Polarity) to understand thetherapeutic efficacy (in traditional terms) of a single or a formulatedmedicine” is done by using an instrumental and software based program.

In yet another embodiment of the present invention, from a largedatabase prepared using this method, it gives many generalizations ofthe therapeutic efficacy of a particular group of plants,therapeutically classified as a group for a particular disease.

In yet another embodiment of the present invention using the X, Y, R, G,B as coordinates of a selected peak in the fingerprint, a barcode isgenerated using a bar coding software, which makes the medicineproprietary for an industry.

In yet another embodiment of the present invention, 3-D chromatogram ofthe medicine is analyzed using all its 3 dimensional properties of thesaid image. If the 3-D chromatogram is considered as a cap with a hood,the matching of the entire cap 3 dimensionally, with another sample ofdifferent qualitative and quantitative properties, the extent it matchedis presented as an analytical report qualitatively and quantitatively.Here the hood of the Cap is compared to the peak of the molecule at aparticular wavelength. A sample with more number will like a cap withmany hoods. Thus the matching of the three dimensional coordinates willprovide a foolproof method of comparison and analysis. The coordinate itmatched will give qualitative and the extent it matched will give thequantitative data of the sample understudy. This is made possible byspecial software prepared for this purpose. This becomes an ultimatemethod of quality control.

In yet another embodiment of the present invention relates to a methodto provide a novel chromatographic finger printing of herbal medicinesand formulations using the contour and 3-D chromatograms of the herbalmedicines and formulations is proposed. They are developed on a PhotoDiode Array detector (PDA) of a High Pressure Liquid Chromatograph. Thisdelineates the data of the spectral properties of the constituentspresent in the herbal medicines presented in a specific order ofpolarity under experimental analytical conditions.

In yet another embodiment of the present invention relates to a to useUV-Visible spectra of the compounds which provides the conjugativeproperties of the molecules and the concentration of the individualconcentrations of the molecules along with the polarity of themolecules.

In yet another embodiment of the present invention relates to a methodto provide the UV_VIS spectra of all the constituents shown in a singleimage “The Chromatographic Fingerprint”. The fingerprint thus becomesthe blue print of the constituents present in a single medicine orformulation for an assay and quick identification of the medicineunderstudy.

In yet another embodiment of the present invention relates to use offingerprint of contour and 3-D chromatograms as a basis for theidentification of chemical constituents to limit the scope of theinvention.

In yet another embodiment of the present invention relates to the methodhaving standard analytical parameters like Extraction with ethylalcohol, maintaining same run time 0-60 min, using same mobile phaseacetonitrile along with phosphate buffer having a pH in the range of5.5-7.5, and a same UV-Visible Range of 200-800 nm.

In yet another embodiment of the present invention, a method uses astandard analytical parameter like Extraction with same solvent ethylalcohol for all samples for the finger printing of a particulartherapeutic group of samples to make therapeutic generalizations.

In yet another embodiment of the present invention relates to a methodof fingerprinting for the adulterated food, drug and chemical samplesand to identify the pure and adulterated.

In yet another embodiment of the present invention relates to a methodof fingerprinting for the substituted food, drug and chemical samplesand to identify the pure and the substituted.

In yet another embodiment of the present invention relates to a methodof fingerprinting for the contradictual food, drug and chemical samplesand to identify the pure and the substituted.

In yet another embodiment of the present invention relates to a methodof fingerprinting for the commercial samples of food and drug and toidentify the pure and the substituted.

In yet another embodiment of the present invention relates to a methodof fingerprinting for the organic and organo-metallic constituents inany type samples to identify the chemical constituents present in it forvarious purposes of quality control and process standardization.

In yet another embodiment of the present invention relates to a methodof fingerprinting for the Allopathic, Ayurvedic, Homoeo, Siddha, Unani,Chinese medicine, Tibetan and Kampo (Japanese) medicine samples for thequality control and chemical and therapeutic standardization

In yet another embodiment of the present invention relates to a methodof fingerprinting for the study of variation of chemical constituents inNaturally occurring samples and to identify and standardize the chemicalconstituents in them.

In yet another embodiment of the present invention relates to a methodof fingerprinting for the study of variation of chemical constituents inNaturally occurring samples and to identify and standardize thevariation in chemical constituents in them due to geological andEcological factors.

In yet another embodiment of the present invention relates to a methodof fingerprinting for the study of variation of chemical constituents inNaturally occurring samples and to identify and standardize thevariation in chemical constituents in them due to Genotypic andPhenotypic variation factors.

In yet another embodiment of the present invention relates to a methodof fingerprinting for the study of chemical constituents inSynthetically prepared samples and to identify and standardize thechemical constituents in them for chemical and therapeuticstandardization which ever is applicable.

In yet another embodiment of the present invention relates to a methodof fingerprinting for the study of chemical constituents in herbalproducts of single and formulated medicine samples and to identify thechemical constituents in them for chemical and therapeuticstandardization.

In yet another embodiment of the present invention relates to a methodof fingerprinting for the study of variation of chemical constituents inbiological samples and to identify and standardize the chemicalconstituents in them.

In yet another embodiment of the present invention relates to a methodof fingerprinting for the study of variation of chemical constituents indifferent brands of products of single and formulated food and medicinesamples and to identify the chemical constituents in them for chemicaland therapeutic standardization.

In yet another embodiment of the present invention relates to a methodof preparing a database giving many generalizations of the therapeuticefficacy of a particular group of plants, classified as a group for aparticular disease or therapeutic classification.

In yet another embodiment of the present invention relates to a methodto develop fingerprinting and to categorize and quantify theconstituents of a medicine, based on polarity and conjugation from 3-Dand contour chromatograms.

In yet another embodiment of the present invention relates to a method,which provides a barcode for the selected peak of a molecule given inthe image, wherein X—the retention time, Y—the wavelength, R—the numberof red pixels, G the number of green pixels and B the number of bluepixels are the coordinates provided by the present computer based(Microchip, Dongle switch, hardware and software locked) software and isfeed to any commercially available re-salable bar-coding softwareresident in the proposed software, generates a barcode. Some examples ofimages displaying the X—retention time, Y—wavelength, R—number of redpixels, G—number of green pixels and B—number of blue pixels thecoordinates of a particular peak (s), which is specific to the productand the barcodes thus, generated are enclosed.

In yet another embodiment of the present invention relates to a method,to prepare a database of barcodes for the fingerprints developed usefulfor all types of database applications.

In yet another embodiment of the present invention relates to a methodto generate display windows for all the samples the fingerprints aredeveloped. In the ‘display window’ all details of the samples like both3-D and Contour fingerprints, the barcode, details of the origin(Industry or Country), manufacturing date, date of expiry, reporteddosha, individual constituents used, their assay, batch number, lotnumber, M.R.P and other information on the label), are displayed. Whenthe barcode on the label is shown to the vending machine, it willdisplay the attached display window. This helps to know the chemical andtherapeutic authenticity of the medicine being sold/purchased in alltypes of regulatory and commercial applications.

In yet another embodiment of the present invention relates to a methodto attach the display windows with the respective barcodes, facilitatingto deal with display windows in all applications whenever they are usedas a source of data and information.

In yet another embodiment of the present invention relates to a methodto prepare a database of display windows thus generated and attached tothe respective barcodes, to use in the ENTERPRISE RESOURCE PLANNING(ERP) and CUSTOMER RELATIONSHIP MANAGEMENT (CRM) applications for allcommercial networking transactions of the medicines and samples forwhich the database was prepared.

In yet another embodiment of the present invention relates to a methodto prepare a database of barcodes and display windows and anyinformation, specially required for the regulatory authorities tocontrol the movement of the medicines in and out of the country.

In yet another embodiment of the present invention relates to a methodfor chromatographic fingerprinting which enables to understand andstandardize the Physico-Chemical properties of the medicines like colorfor the therapeutic standardization of the medicines and humors.

In yet another embodiment of the present invention relates to a methodfor chromatographic fingerprinting which enables to understand andstandardize the Physico-Chemical properties like Taste (Rasa) like Sour(Amla), Salty (Lavana), Pungent (Katu), Bitter (Tikta), Astringent(Kashaya) of the medicines, for the therapeutic standardization of themedicines and humors.

In yet another embodiment of the present invention relates to a methodfor chromatographic fingerprinting which enables to understand andstandardize the Chemical properties of the medicines like Cold, Hot,Slow in action, Sharp in action, Heavy, Light, Soft, Lubricated, Supple,Dry (Guna's like Sheeta, Ushna, Manda, Teekshna, Guru, Laghu, Snigdhaand Rooksha as described in Ayurveda) of the medicines for thetherapeutic standardization of the medicines.

In yet another embodiment of the present invention relates to a methodfor chromatographic fingerprinting which enables to understand andstandardize the Physico-Chemical properties of the medicines likePotency, Metabolite formation after assimilation and Specific propertieslike Charlotte of the molecules (Described as Veerya, Vipaka andPrabhava) for the therapeutic standardization of the medicines andhumors.

In yet another embodiment of the present invention provides a softwarebased data processor of 3-D chromatograms and contour image of aningredient.

The method of the invention is described in steps with reference to theaccompanying tables, drawings, flow charts and examples, which areprovided to illustrate some of the embodiments of the invention, and thesame should not be construed as limitations on the inventive conceptembodied herein.

DESCRIPTION OF THE ACCOMPANYING TABLES AND FIGURES

The following examples are given by way of illustrations and theseshould not be construed to limit the scope of the invention.

I. TABLES

-   1. Table showing different philosophies and various terminology used    in different medicines-   2. Table showing Relation of Humors, Properties, and different parts    of the human body—An Ayurvedic approach.-   3. Table showing Division in terms of the Macrocosm in Chinese    medicine-   4. Table showing Division in terms of the Microcosm in Chinese    medicine-   5. Table showing the relation of five natural elements and their    relation-   6. Table showing the meaning of Yin and Yang used in Chinese    Medicine-   7. Table showing the basis of color for the therapeutic    classification of the medicines-   8. Table showing the effect of different colors on different    diseases-   9. Table showing Properties of the six tastes (Rasas in Ayurveda)    and their properties and efficacy-   10. Table of Colors and The Relation with Wavelengths.-   11. Table showing the role of acidity and alkalinity in human body-   12. Table showing the comparison of different analytical techniques    being used for the finger printing and chemical standardization.-   13. Table showing the parameters used for developing fingerprints of    some of the medicines-   14. Table showing the therapeutic classification of medicines    reported in the proposed invention.-   15. Table of medicines shown as thumbnails.-   16. Table showing the division of the fingerprint in to therapeutic    zone based on the conjugation and polarity.

II. FIGURES

FIGS. 1A & B shows the five basic elements in Chinese medicine and therelationship between them. Imbalance (excess or deficiency) of any oneelement leads to disturbance in other elements and becomes the rootcause of a disease. The health of human body is achieved by managing andcontrolling the above eleiments in Chinese medicine.

FIG. 2 shows the effect of colors on the basic humors based on which themedicines of the same colors were selected for vitiating thecorresponding humor. The color of a medicine is due to the chemicalproperties of the constituents present in it, and thus indirectly thechemical properties are used for the therapeutic standardization.

FIG. 3 shows the fingerprints of Shilajit of two different brands. Thechemical profile in the fingerprint shows the therapeutic efficacy dueto the presence of more number of molecules with wide conjugativeproperties. The chemical profile varies with the age of the sample, itspent in the earth, and the more it is old, the more it will betherapeutically active and may depend on the place of collection andpurification process.

FIG. 4 shows the existing use of chromatographic fingerprints on alabel.

FIG. 5 shows chromatogram of Commercial Turmeric (Food).

FIG. 6 shows chromatogram of Furazolidine (Allopathic).

FIG. 7 shows chromatogram of Krimikutara Ras (formulation).

FIG. 8 shows chromatogram of Shilajit (good by efficacy).

FIG. 9 shows chromatogram of Shilajit (Poor by efficacy).

FIG. 10 shows chromatogram of Suryavarti (formulation).

FIG. 11 shows chromatogram of Tea (food).

FIG. 12 shows chromatogram of Trikatu (formulation).

FIG. 13 shows the fingerprints of all yellow medicines. Here thefingerprint of Sandigdha Dravyas (a controversial drug) shows a cleardifference in appearance, making the identification more easy.

FIG. 14 shows the fingerprints of all medicines of PITTA HARA in nature.The presence of constituents in zone-1 indicates the efficacy of themedicine.

FIG. 15 shows the fingerprints of all medicines of KAPHA HARA in nature.The presence of constituents in zone-2 indicates the efficacy of themedicine.

FIG. 16 shows the fingerprints of all medicines of VATA HARA in nature.The presence of constituents in zone-3 indicates the efficacy of themedicine.

FIG. 17 shows the fingerprints of all medicines of PITTA KAPHA HARA innature. The presence of constituents in zone-1 and zone-2 indicates theefficacy of the medicine.

FIG. 18 shows the fingerprints of all medicines of KAPHA VATA HARA innature. The presence of constituents in zone-2 and zone-3 indicates theefficacy of the medicine.

FIG. 19 shows the fingerprints of all medicines of PITTA VATA HARA innature. The presence of constituents in zone-1 and zone-3 indicates theefficacy of the medicine.

FIG. 20 shows the fingerprints of all medicines of TRI DOSHA HARA innature. The presence of constituents in all three zones indicates theefficacy of the medicine.

FIG. 21 shows the fingerprints of Kali musali and Safed musali that areused as tri-doshahara medicine.

FIG. 22 shows fingerprints of different samples of CitrallusColosynthis. The fingerprint shows the lack of some constituents due towhich this method is used for standardization of extraction process ofhomoeo mother tinctures from plants.

FIG. 23 shows fingerprints of different samples of HolarrenaAntidyssentric collected from different places of the country. Thefingerprint shows the influence of ecological factors on the chemicalconstituent of the plant material.

FIG. 24 shows the fingerprints of two samples of Beetle leaves fromdifferent places. The flavonoids present in the time range of 30-40 minshows the influence of genotypic, phenotypic variations and ecologicalfactors on the chemical constituents of the plant material.

FIG. 25 shows the satellite images of India. These satellite imagesindicates that India has different tropical zones.

FIG. 26 shows the fingerprints of two formulations used as cosmeticslike Herbal head Bath powders.

FIG. 27 shows the fingerprints of TRIKATU of two different brands. Thedifference in their assay may be due to variations in the constituentelements of TRIKATU.

FIG. 28 shows the fingerprints of turmeric and its three differentcommercial products. A common peak occurs at 20 minutes in all thesefingerprints.

(FIGS. 29 to 92 shows the finger prints of all the medicines reported inTable 13)

FIG. 29 shows both fingerprints of whole plant of Abel moschus,Moschatus medicum.

FIG. 30 shows both fingerprints of bark of Acacia suma.

FIG. 31 shows both fingerprints of leaflets of Acalypha indica.

FIG. 32 shows both fingerprints of leaves of Adhatoda vasaka

FIG. 33 shows both fingerprints of leaves of Adiantum caudatum

FIG. 34 shows both fingerprints of stem bark of Ailanthus excelsa

FIG. 35 shows both fingerprints of rhizome of Acorus calamus

FIG. 36 shows both fingerprints of big single cloves of Allium porum

FIG. 37 shows both fingerprints of small cloves of Allium sativam

FIG. 38 shows both fingerprints of rhizome of Alpinia galanga

FIG. 39 shows both fingerprints of rhizome of Alpinia officinarum

FIG. 40 shows both fingerprints of rhizome of Alpinia speciosa

FIG. 41 shows both fingerprints of unprocessed raw fruit nut of Arecacatechu

FIG. 42 shows both fingerprints of milk processed nut of Areca catechu

FIG. 43 shows both fingerprints of stem bark of Areca kateeh

FIG. 44 shows both fingerprints of Homoeo mother tincture of Arnica

FIG. 45 shows both fingerprints of whole herb of Bacopa monneri

FIG. 46 shows both fingerprints of stem bark of Berberis aristata

FIG. 47 shows both fingerprints of whole plant of Borrhievia diffusa

FIG. 48 shows both fingerprints of big, ripped fruit of Capscicum Annumlinn.

FIG. 49 shows both fingerprints of big unriped, fruit of Capscicum annumlinn.

FIG. 50 shows both fingerprints of small, unriped, fruit of Capscicumannum linn.

FIG. 51 shows both fingerprints of stem bark of Coscinium fenestratum

FIG. 52 shows both fingerprints of root and leaf of Coccinidium grandis

FIG. 53 shows both fingerprints of leaf of D actlylactinium Aegyptium(erect)

FIG. 54 shows both fingerprints of leaf of Dactlylactinium Aegyptium(prostrate)

FIG. 55 shows both fingerprints of leaf and bark of Diristachiscineraria

FIG. 56 shows both fingerprints of fruit epicarp of Emblica officinalis

FIG. 57 shows both fingerprints of formulation of a face pack

FIG. 58 shows both fingerprints of formulation of a face pack

FIG. 59 shows both fingerprints of root bark of Glycerrhzia glabra

FIG. 60 shows both fingerprints of powder of whole plant of Glycerrhziaglabra

FIG. 61 shows both fingerprints of whole plant of Gymnema sylvestrae

FIG. 62 shows both fingerprints of stem bark of Hollerona Antidysentrica

FIG. 63 shows both fingerprints of root of Innula recemosa

FIG. 64 shows both fingerprints of flower of Michellia champaka

FIG. 65 shows both fingerprints of Leaf of Moringa olifera

FIG. 66 shows both fingerprints of homeopathic mother tincture of Myricacerefera

FIG. 67 shows both fingerprints of whole plant of Nahi axillae

FIG. 68 shows both fingerprints of stem bark of Oroxylum indicum

FIG. 69 shows both fingerprints of leaf of Ocimum sanctum

FIG. 70 shows both fingerprints of leaf of Pluchea lanceolata

FIG. 71 shows both fingerprints of stem bark of Picrorrhiza kurroh

FIG. 72 shows both fingerprints of leaf of Piper beetle.

FIG. 73 shows both fingerprints of seeds of Psoralia corilifolia

FIG. 74 shows both fingerprints of leaf of Raphanus sativus

FIG. 75 shows both fingerprints of root of Ricinus cummunis

FIG. 76 shows both fingerprints of stem and root of Rubia cordifolia

FIG. 77 shows both fingerprints of root of Saussrea lappa

FIG. 78 shows both fingerprints of whole herb of Spheranthus indicus

FIG. 79 shows both fingerprints of stem bark of Symplocus racemosus

FIG. 80 shows both fingerprints of fruit of Terminalia chebula

FIG. 81 shows both fingerprints of fruit of Terminalia bellerica

FIG. 82 shows both fingerprints of whole plant Trigonella faenum g.

FIG. 83 shows both fingerprints of stem and root of Tribulusterrestrias.

FIG. 84 shows both fingerprints of leaves of Tylophora asthmatica

FIG. 85 shows both fingerprints of mother tincture of Homoeo medicineViburnum

FIG. 86 shows both fingerprints of Root of Withinia somnifera

FIG. 87 shows both fingerprints of rhizome of processed Zinziberofficinalis

FIG. 88 shows both fingerprints of powder of Avipattakara churna

FIG. 89 shows both fingerprints of an herbal formulation of Kamaduga Ras

FIG. 90 shows both fingerprints of a Kumarayasava a herbal medicine byfermentation process

FIG. 91 shows both fingerprints of an herbal formulation of Mahalakshmivilas ras

FIG. 92 shows both fingerprints of an herbal formulation of Suvarnayogaraja Guggulu

FIG. 93 shows the fingerprint of Anandabhairavi Ras. Right clicking onany particular peak, the image software will display the X Y, R, G and Bcoordinates of the peak, which are used for bar coding. Thesecoordinates are shown inside a box (near the peak) and in the tool bar.

FIG. 94 shows the fingerprint of Krimikutara Ras. The image softwaredisplays the bar code values of a particular peak.

FIG. 95 shows the barcode generated for Anandabhairavi Ras

FIG. 96 shows the barcode generated for Krimikutara Ras.

FIG. 97 shows the display window for Anandabhairavi Ras

FIG. 98 shows the display window for Krimikutara Ras

FIG. 99 shows how the network works in an Enterprise Resource Planningand Customer Relationship Management applications networked using thedatabase prepared by the proposed method.

FIG. 100 shows a new chromatogram in the form of a colored bar chart forthe tender leaves of Azadiracta indica collected in February

FIG. 101 shows a new chromatogram in the form of a colored bar chart forthe medicine of Anandabhairavi Ras an herbal formulation.

FIG. 102 shows a new chromatogram in the form of a colored bar chart forthe medicine of Krimikutara Ras, an herbal formulation.

FIG. 103 shows the Pie diagram for the chromatogram of Azadiracta indica

FIG. 104 shows the Pie diagram for the chromatogram of AnandabhairaviRas

FIG. 105 shows the Pie diagram for the chromatogram of Krimikutara Ras

FIG. 106 shows the Pie diagram of vitiation disorders (dosha)quantitative.

FIG. 107 shows the Pie diagram of vitiation disorders (dosha)quantitative.

FIG. 108 shows the Pie diagram of vitiation disorders (dosha)quantitative.

FIG. 109 shows the fingerprints as 3D and contour images of severalherbal medicines, which are fed into a database and are used for variousERP and CRM applications.

FIG. 110 shows the fingerprints as 3D and contour images of severalherbal medicines, which are fed into a database and are used for variousERP and CRM applications.

FIG. 111 shows the fingerprint of adulterated cosmetic samples as 3D andcontour images.

FIG. 112 shows the fingerprints of Mother tincture as 3D and contourimages these can be used to find out the dilution of Mother tincture.

FIG. 113 shows the fingerprints of isolated medicines and their UVspectra as 3D and contour images.

FIG. 114 shows the fingerprints of allopathic medicines as 3D andcontour images.

FIG. 115 shows a diagram with operational sequence of the software withvarious Functionality's.

The invention is described in detail below with reference to theaccompanying drawings, flow charts and examples, which are provided toillustrate some of the embodiments of the invention, and the same shouldnot be construed as limitations on the inventive concept embodiedherein.

Method of Chemical Standardization:

Hence unlike a method currently under use, where in a chromatogram isgiven at a single wavelength, a novel method of chromatographicstandardization, finger printing and bar coding is proposed, usingcontour and 3-D chromatograms. It provides the total chemical profile(properties like polarity and conjugation, there in) of the chemicalconstituents present in complex medicines like herbal medicines andformulations or any medicine. Further, bar coding the finger prints thusgenerated will provide many commercial features in dealing suchmedicines using the ERP and CRM applications.

The existing method of TLC fingerprinting FIG. 4 being used as achromatographic finger print, is showing only an assay of theconstituents present in it. It is not providing any chemical propertylike conjugation or polarity. Another method of fingerprinting by HPLCshows a chromatogram at a single wavelength presented as a“Chromatographic Finger Print” of the medicine. In this, a selected peakis identified chemically, what it is by structure, using various otheranalytical techniques like NMR, LC-MS and IR for structural elucidation.So, the single chromatogram by it self is not able to say what theefficacy of the medicine is, without the support of other costlieranalytical instruments. It will be highly impractical to use such costlytechniques for a complex herbal medicine and formulations prepared byformulating various organic and inorganic medicines for a particulartherapeutic purpose.

The quality of any formulated medicine will depend on the process withwhich it was made. This will be different for each pharmacy orpharmacist. What actually needed for the quality control of herbalmedicines and formulations is a simple analytical method that can givethe number of constituents (qualitative and quantitative) present in asingle medicine or formulation, and the therapeutic efficacy of themedicine under study. Hence, any method, which does not provide theabove information, is incomplete.

In the method of chemical standardization of the present invention, theconstituents were first extracted in to a suitable solvent. The extractwas subjected to separation into individual constituents on a HighPressure Liquid Chromatograph under standardized analytical conditions.The 3-D and contour chromatograms given by the instrument were convertedin to chromatographic fingerprint images. The images were analyzed usingimage analysis software specially prepared for this work. The out putdata is interpreted for the said standardization. Detailed descriptionof the method is given in experimental description of the method.

Method of Therapeutic Standardization:

The traditional therapeutic standardization is highly individualistic byability and perception of the doctor. A general availability of suchmethod will be practically difficult. But the existing scientificscenario emphasizes that any method or mechanism needs to bestandardized, and reproducible. Hence, in the present method of chemicaland therapeutic standardization an instrumental method is proposed whichbrings down the human factor. This is made possible by an instrumentalanalytical technique, which explains about the chemical and therapeuticefficacy of the medicines under study in a simple way. In a mostscientific and organized society of modern science, the knowledge ofassessing the therapeutic efficacy of the medicines should be explainedwith rational justification rather than individual skills and abilities,as they will differ from individual to individual and arenon-reproducible. The method of the invention envisages the same withoutdeviating from the traditional concepts.

As explained above if one can assess the therapeutic efficacy of themedicine by the physico-chemical properties (Polarity and conjugation),the activity of the medicines is understood thus achieving thetherapeutic standardization. In the present method, the conjugative andpolarity properties are taken in to consideration to assess thetherapeutic efficacy of a medicine.

In the ancient literature a clear classification of soils and plantswere given based on their physico-chemical nature and therapeuticefficacy. The selection of medicines for a particular disease was donebased on the guidelines like color, texture, odor and physicalappearance. Table 8 of the effect of different colors on different bodyparts shows how color was used for this purpose. The soil types and thediversity of the drug action were also mentioned while selecting amedicine. The effect of climate and its effect in the efficacy on thedrug plants were also clearly mentioned. Because the chemicalconstituents present in the plant depends on these geological andecological variable factors, guide lines were laid down for the place ofcollection, time (seasonal and daily) of collection, part of plant forcollection and age of plant for collection, required for a specifictherapeutic action.

Based on the generalia of the plants used for a common type of diseasesthe plants were classified into 37 groups (Wealth of Susrutha, K. H.Krishnamurthy, Indian Institute of Ayurveda, Coimbatore,India—originally from Susruta Samhita sutrasthana 38). Thus, theseplants should contain the chemical constituents having similartherapeutic efficacy on the reported disease.

When the fingerprints of the different classes were studied, some commonfeatures are found about the therapeutic efficacies of the medicines.The same efficacy was reported in the traditional literature also. Inother words, i.e. the experimental and reported results are equal.Hence, studying different medicines, having different therapeuticefficacy validated the method.

The FIG. 13 shows all medicines, which are yellow in color. In theancient literature of Ayurveda all these medicines were classified asHaridra class, all the medicines being yellow in color like Haridra(Turmeric). When the fingerprints are studied, it is found that allthese medicines are reported to be used as Kapha Hara, vitiation ofdisorders related to mucogeneous constituents of the body. Hence, it isunderstood that the color of the medicines has a direct relation withtheir therapeutic efficacy. Rationally it is also true that the colorand the efficacy of the medicines are due to chemical constituents andtheir physico-chemical properties.

The fingerprints of single medicines like Rubia cordifolia, SaracaIndica, Picrorrhiza Kurro, Phyllanthus Niruri and Formulations likeArogya Vardhini and Avipattakara Churna are presented in FIG. 14. Themolecules eluted in Zone 1 indicates the presence of Polar constituentsbased on the elution pattern due the set analytical conditions. Ageneral tendency of this elution pattern for the medicines reported tobe Pitta Hara, confirms that high polar constituents act mainly as PittaHara.

The fingerprints of single medicines like Zinziber officinalis(Processed), innula racimosa, Sausserea Lappa, Ocimum Sanctum, Glycerziaglabra and Shilajit are reported to be Kapha Hara. The molecules elutedin Zone 2 indicate the presence of Medium Polar constituents. A generaltendency of this elution pattern for the medicines reported to be KaphaHara confirms that medium polar constituents act mainly as Kapha Hara asshown in FIG. 15.

The fingerprints of single medicines like Alpinia offinarum, Ricinuscommunis, and Formulations like Suvarna yogaraja Guggulu, Brihatvatachintamani with swarnamakshkam, Huthasana and Mahayogaraja Guggulu arepresented in this Figure. The molecules eluted in Zone 3 indicate thepresence of very low or non-polar constituents, mostly oily in nature,based on the elution pattern due the set analytical conditions. It isobserved that any medicine used for this disorder contain or mixed withoils. Along with oily type of constituents, the herbo mineral organometallic molecules eluting in this zone are also found Vata Hara. Ageneral tendency of this elution pattern for the medicines reported tobe Vata Hara, confirms that low or non-polar constituents act mainly asVata Hara as shown in FIG. 16.

The fingerprints of single medicines like Azadiracta indica, Curcumalonga, Hollarrheana Antidyssentrica, Berberis aristata, PsoraliaCordifolia and Citrullus Colosynthis are presented in this Figure. Themolecules eluted in Zone 1 and 2 indicate the presence of high polar andmedium polar constituents. Thus, these medicines having the medium polarconstituents are found to be Pitta-Kapha Hara. This confirms that theefficacies of the medicines are under stood by the polarity of theconstituents present in it as shown in FIG. 17.

The finger prints of the single medicines like Tribulus Terrestrius,Moringa Olifera, Piper Beetle and formulations like Trikatu indicate thepresence of constituents in the Zone 2 and Zone 3 indicating theefficacy as Kapha Vata Hara in nature, FIG. 18 explain the same.

The fingerprints of the single medicines like Bacopa monneri, OroxylumIndicum and formulations like Kanchanara Guggulu indicate the presenceof constituents in the Zone 1 and Zone 3 indicating the efficacy asPitta Vata Hara in nature. In a formulation called Anadabhairavi, eventhough the efficacy reported is Pitta Vata Hara it is found Kapha VataHara by finger printing. This indicates the artificial preparation ofthe medicine was not successful to prepare a medicine of the requiredefficacy. Hence, this method is useful in the process standardization ofthe preparation of complicated formulations mentioned above. FIG. 19indicate the same.

The finger prints of the single medicines like Allium Cepa, WithiniaPubiscence (Red Seeds), Embalika Officinalis and formulation likeMahalakshmi vilas ras indicate the presence of constituents in all thethree Zones of 1,2 and 3 indicating the presence of molecules of theentire range of polarity. This indicates that they will be of Tri DoshaHara by efficacy. In the finger print of Mahalakshmi vilas ras thepresence of two similar type of molecules can be seen like isomeric innature. The Prabhava effect is understood in these type of medicineswhen such type of isomeric (Geometrical and Chiral) constituents arepresent. FIG. 20 shows the fingerprints of all medicines of Tri DoshaHara.

The fingerprints of Kalimusali (Curculigo Orchioidis) and Safedmusali(Asparagus Adescendens) indicate how two plants of different family'swere classified under same therapeutic group. The fingerprints showsimilar constituents in all the three zones with little difference inassay indicating the tri-doshahara property, as indicated in FIG. 21.

The fingerprints of a single medicine of two different sources likeCitrullus Colosynthis used in both Ayurveda and Homoeo are given. Onobservation of the fingerprints, it is found to contain constituents ofthree polarities, but mostly high polar molecules more in number. Oncareful observation of the fingerprints, it is observed that thepresence and absence of molecule at 12 minutes is the only differencebetween both the images. The taste of first medicine was very bitterwhen compared to the second one. Thus using the taste, as a measure ofthe efficacy of the medicines is also proposed, this was mostly used inthe ancient literature, as shown in FIG. 22.

The fingerprints of Hollarrena Antidysentrica, a medicine collected fromtwo different parts of the country has shown much difference in theirchemical profile. This indicates the influence of geological,ecological, genotype and phenotypic and other variable factors on thechemical constituents of the herbal medicines; this is illustrated byFIG. 23.

A vast difference was seen in two samples of beetle leaves one fromAndhra Pradesh and another from Calcutta of India FIG. 24. This confirmsthe role of ecological, genotypic and phenotypic variations in thechemical constituents of plant parts.

In the FIG. 25 the Eco-regions, precipitation, Temperature and Climateof India were shown to understand the role of seasons on the ecology ofthe flora and fauna of it. The variations in the seasons will have animpact on the chemical constituents of the herbal plants and therebymedicines produced from them. This applies to the entire world wheneveran herbal plant is collected from different parts of the world.

The fingerprints of two formulations used as cosmetics like Herbal headBath powders are given. The fingerprint of the pure herbal material istotally different from the adulterated one. The artificial detergentsand foaming agents eluted at 25 to 40 minutes are clearly seen in theadulterated sample, they are highly basic and soapy in nature. Thissupports that the method of the invention is useful for the regulatoryauthorities to check the pilferage of traditional medicines withadulterations and substitutions, as illustrated in FIG. 26.

The fingerprints of a formulation namely Trikatu of two different brandsshows difference in its assay. This may be due to usage of singlemedicine(s) were used to prepare the formulations from differentsources. The present method shows the extent they are differentqualitatively and quantitatively, facilitating to prepare standardizedmedicines and extracts of herbal medicines as shown in FIG. 27.

The fingerprints of a single medicine of three different brands of thesame food material like turmeric is given in FIG. 28. In the fingerprint of the natural turmeric it is observed that the yellow curcuminmolecules eluted at 20 minutes. The same molecules are seen in all thebrands commonly. The difference in the profile is the commercial samplesare because they were prepared processed (Boiled) turmeric rhizomes andthe natural is from unprocessed (Un-boiled) rhizomes.

FIGS. 29 to 92 gives the fingerprints developed for various medicinesand the image parameters (Elevation and rotation) used for the 3-D andContour chromatograms of all the medicines given in table 13. In tableno 14, the medicines analyzed were classified on the basis oftherapeutic efficacy. The analysis of fingerprints of the respectivemedicines using the proposed software will support the claim of utilityof the method of the invention for the therapeutic standardization.

This confirms that this method is useful in many purposes of dealing thetraditional medicines. It is useful for modern medicines also tounderstand their therapeutic efficacy in traditional terms.

Method of Bar Coding, Enterprise Resource Planning (ERP) and CustomerRelationship Management Application (CRM) Application

In the present method the software analyzes the image and can displaythe coordinates X retention time, Y wavelength, R number of red pixels,G number of green pixels and B number of blue pixels by the presentcomputer based (Microchip, Dongle switch, hardware and software locked)software for a contour chromatogram, for a particular peak which isspecific to the product. When this data is transferred/feed to aresident in built re-salable bar-coding software, automatically anoptional pixel value of a peak in the image, it generates the barcodehaving attached the ‘display window’ with all details of the productunder study.

The method of the invention facilitates to barcode any number ofconstituents present in a chromatographic finger print of a herbalmedicine, using the X retention time, Y wavelength, R number of redpixels, G number of green pixels and B number of blue pixels as thecoordinates provided by the present computer based (Microchip, Dongleswitch, hardware and software locked) software and the values of each ofthe constituents provided by the present software. These factors will berepresenting the chemical and therapeutic efficacies of theconstituents. Hence instead of a mere catalogue number presently usedfor the bar coding, a novel method of bar coding is proposed where in,the coordinate values of the constituent will be displayed along withother details. Thus, the barcode and the coordinates-will speak aboutthe chemical and therapeutic properties of the product.

It becomes a tool for the regulatory authorities like Drug Controller,Public analysts, Food adulteration enforcement authorities, Forensic andCustoms and Central excise departments for the regulation of the herbalProducts. The fingerprint of the medicines should be printed on thelabel and should be tallied when it is checked. This also helps tomonitor the various other brands of a same medicine for an industry tomonitor. FIGS. 93-94 shows how the software gives the coordinates for aselected peak of the image. These values will be given for a barcode tobe generated. FIGS. 95, 96 are the barcodes thus generated. FIGS. 97, 98show how a display window will be for an herbal medicine with all labeldetails. The barcode will be able to show when the display windows whenthey are attached to the barcodes of the respective products. When alarge database is prepared for the products and made available in thenetwork, any ERP and CRM application can use for any required purpose bynetwork. FIG. 99 shows how the network woks in an ERP and CRMapplications networked.

Various Steps Involved in the Present Invention

In the present method of analysis a Validated High Pressure LiquidChromatograph equipped with a Binary Gradient system of pumps, a PhotoDiode Array Detector (PDA), and a Software based data processor forpresentation of the chromatograms was used. After the complete elutionof all ingredients, the 3D and contour chromatograms (having theinformation of the UV-Visible Spectra, absorbance and retention times ofall the constituents present in a single medicine or formulation) wereconverted into an image and proposed as a finger print. This enjoys themerit of not requiring any internal or external standard sample for anauthentic qualitative and quantitative analysis of all the ingredientspresent in a medicine, unlike in the present method of analysis ofmedicines.

Experimental Description of the Method:

The proposed method is described in 4 steps with reference to theaccompanying drawings, flow charts and examples, which are provided toillustrate some of the embodiments of the invention, and the same shouldnot be construed as limitations on the inventive concept embodiedherein.

The entire method is described in the steps mentioned below:

-   Step 1: Selection of medicines and extraction of the constituents-   Step 2: Separation of the constituents into individual constituents    and generating and converting the 3-D and Contour Chromatograms in    to finger prints.-   Step 3: Analysis of the fingerprints using the software developed.-   Step 4: Interpretation of data    Description of the Present Method of Analysis    Step 1: Sample Preparation

Constituents are extracted from the medicines using ethyl alcohol,selected based on the chemical nature (polarity) of the sample. When thepH of the aqueous alcohol extract is varied, the extraction ofconstituents also has varied. The basic pH has extracted more number ofconstituents than acidic pH. Suitable pH was selected for extraction ofdifferent medicines and the same was maintained using buffers. The roleof acidity and alkalinity was taken into consideration while selectingthe pH for extraction.

Step 2

Experimental Work Done on the Instrument:

The extract was subjected to separation analysis, using High-PressureLiquid Chromatographic (HPLC) instrument. In the present method ofanalysis, a Validated High Pressure Liquid Chromatograph equipped with aBinary Gradient system of pumps, a Photo Diode Array Detector (PDA), anda Software based data processor, for the preparation of thechromatograms were used. A known amount of the sample (say 20 ul) ofextract is injected into rheodyne injector (fitted with 20 ul loop).Elution of the sample was performed with suitable time programmedgradient system of mobile phase at a fixed flow (1 ml/min). Care istaken that no part of the sample is left in the column un-eluted. Thefollowing analytical conditions set for the analysis.

-   a. A reverse phase column was used along with a time programmed    gradient elution of an aqueous phosphate buffer (In the pH range of    5.5-7.5) and a non-aqueous solvent (acetonitrile or methanol) is    used as eluent based on the chemical nature of the sample under    analysis.-   b. A wave length range of 200 to 800 nm was used for the PDA    detector and the run time is fixed based on the time program.-   c. The time program, which changes the concentration of non-aqueous    solvent like Acetonitrile 0-100% of organic solvent, is used in the    instrumental parameters existing in the instrument.

The instrument was triggered for the analysis after injecting the sampleinto the injector. The run was stopped whenever the analysis iscompleted or the instrument will stop the run automatically after theentire time program is completed.

In the three types of display of data, the chromatograms, one windowdisplays chromatogram at a selected wavelength, in another it displaysthe contour chromatogram which displayed the retention time (run time)of the analysis on X-axis and the wavelength range on Y-axis. In anotherwindow, it displayed the 3-D chromatogram of the sample where in itdisplayed the retention time (run time) of the analysis on X-axis, theconcentration range on Y-axis and the wavelength range on Z-axis. The3-D and contour chromatogram thus developed by the system was convertedinto an image.

The images thus generated were analyzed by the proposed software, whichprovides a novel chromatogram and the qualitative and quantitativeanalytical data of the in-gradients present in the medicines. The pixelvalues represented by different colors from Violet, Indigo, Blue, Green,Yellow, Orange and Red attributed as a measure of the concentration(quantitative) of the constituents proportional to the color. Extractingthe individual colors mentioned above and shows in separate widows foreach color. This is the basis of chemical standardization. Somechromatograms thus generated are shown in FIGS. 100-102.

The chromatogram thus provided by the software gives the information ofconjugative (shown by the UV-VIS absorbance) and polarity properties ofthe individual constituents together. The image is divided into threezones representing, Zone 1 (High polar zone), Zone 2 (medium polar zone)and Zone 3 (low or non polar zone) scaled by retention times based onthe elution pattern, depending on the column used and the mobile phase.Reversing the analytical conditions can reverse the elution pattern.

The 3-D chromatograms of the medicine are analyzed using all its threedimensional properties of the said image. If the 3-D chromatogram isconsidered as a cap with a hood the matching of the entire cap 3dimensionally, with another sample of different qualitative andquantitative properties, the extent it matched will be presented as ananalytical report qualitatively and quantitatively. Here the hood of thecap is compared to the peak of the molecule at a particular wavelength.A sample with more number looks like a cap with many hoods. Thus thematching of the three dimensional coordinates provides a foolproofmethod of comparison and analysis. The coordinate it matched givesqualitative and the extent it matched gives the quantitative data of thesample understudy. This is made possible by special software preparedfor this purpose. This becomes an ultimate method of quality control.

The interaction of the polarity of the molecules being separated, thepolarity of the stationary phase used and the polarity of the mobilephase used for the elution of the sample controls the elution pattern ofthe molecules. The resultant inter action of all the three and otherrelated parameters like temperature etc., decides the elution patternand order of elution of the constituents based on their polarity. Thus,in a medicine all the polar molecules will elute in first ‘Zone 1’(Polar zone of the image), all the medium polar molecules will elute in‘Zone 2’ (Medium polar zone of the image) and all the low polar or nonpolar molecules will elute in ‘Zone 3’ (Non polar zone of the image).When the molecules eluted in these three zones of many fingerprints,many generalizations were made regarding the chemical and therapeuticefficacy of the medicines. This is another basis of therapeuticstandardization. The zones are shown marked in the FIGS. 103-105.

Thus, the chromatogram gives the information, how it is going to actchemically and so therapeutically. When the individual constituentspresent in each zone and represented graphically or by any means of datapresentation, the total constituents of the respective zone gives thepercentage it is going to act on the particular dosha. Thus, the dataexplains how it (medicine) is going to act therapeutically on thevitiation of each dosha collectively based on the qualitative andquantitative properties of the constituents present in the medicine. Forexample if the medicines has 30% constituents in high polar zone (thepixel quantities of various colors like green, yellow, orange and red ofa specific zone as quantities) 70% in medium polar zone it can berepresented as a medicine acts 30% on pitta and 70% on kapha, as thecolors represent different concentrations in the fingerprints. Hence amedicine can be assessed as of Pitta-Kapha Hara (30-70%). Thus, thevitiation of doshas is quantified. This helps the doctor to under standthe efficacy of the medicines and decide his dosage. Some example Piediagrams are given in the FIGS. 106-108.

3-D and contour Spectra of the reported herbal medicines were developedusing the reported analytical conditions. The thumb nail view of themedicines shows how the finger prints can be handled by a software as itis done in the software used in handling the human fingerprints. All thefeatures like searching the similar and compare the similar fingerprintsetc., can be done by inserting the necessary software features. In FIGS.109-114 the thumbnails of the fingerprints for various medicines aregiven. The lists of medicines shown as fingerprints were shown in table15.

Step 3

Analysis of the Image Using Image Analysis Software:

After the complete elution of all ingredients, the 3D and contourchromatograms were converted into images and proposed as fingerprints.This enjoys the merit of not requiring any internal or external standardsample for an authentic qualitative and quantitative analysis of all theingredients present in a herbal medicine, unlike in the analysis of asynthetic medicines.

After developing the image of the 3-D and contour chromatograms of themedicine under study, (Hence forth called as Chromatographic fingerprint) it is analyzed by the soft ware proposed for the analysis ofvarious colors representing the Qualitative and Quantitative propertiesof the constituents present in it.

Scientifically, an image cannot become an analytical data, hence acomputer based image analysis software (software and hard wareprotected) has been developed to analyze the image and give proportionalconcentrations of the ingredients of the medicine under study. Based onthe colors of the constituents present in various retention times andpixel values of the image.

Now the images of the fingerprints were given to Image Analysis softwareas said above. The analysis of various colors was done by which theconstituents will be represented as peaks of the chromatogram and thusproviding a novel presentation of chromatogram in the form of a coloredbar chart. It shows the number of compounds and their conjugativeproperties UV-VIS absorptive property of all of the constituents eluted.The detailed description of the process involved in the analysis of theimage is discussed in the technical features of the software.

The bar chart type of chromatogram thus developed gives a chromatogramhaving a scale of Retention time (0-60) on the X-axis and wavelength inthe range of 200-800 nm, on the Y-axis. It gives the number of pixelsoccupied by each of the colors of each in-gradient in the image,facilitating the qualitative and quantitative analysis of the individualconstituents present in it. Thus, the chromatogram generated ispresenting the number of constituents present in a medicine and their UVabsorption range with quantity of pixels proportional to theconcentration of the molecules.

When the image is divided into, three zones based on the elution patternof the molecules and the changing polarity of the mobile phase. The Zone1 is polar zone as the column used is a reverse phase column, the Zone 2is medium polar zone where in the medium polar molecules are eluted andfinally the Zone 3 is low or non polar zone as the non polar and verylow polar molecules will elute in this zone. Thus, the molecules elutedin zone 1 will be polar, the molecules eluted in the zone 2 will be ofmedium polar in nature and the molecules eluted in the zone 3 will be ofvery low or non polar in nature. Hence, the three zones of the imageswill give the polarity of all the constituents eluted.

Based on the polarity of the molecules eluted, the medicines areclassified according to traditional system of therapeutic efficacy wherein the polar compounds are found to be Pitta Hara, the medium polarcompounds are Kapha Hara and the low or non polar compounds are VataHara. This is the basis of therapeutic standardization of the medicines.The polarity of the constituents is compared to a continuous spectrum ofradiation, where in the dosha is classified as acute to chronic of eachdosha. The starting of the zone will be acute and the end of the zonewill represent the chronic. Thus, the compounds present in the said zonewill act on the said intensity of the disease.

Table 16 shows division of the fingerprint in to different therapeuticzone based on the color of absorption and polarity. The scale on X-axisshows the scale of polarity of the molecules based on the polarity ofthe mobile phase and Y-axis shows the range of wavelength (200-800 nm)absorbed. Based on the reported therapeutic efficacy in the literaturebased on the physico-chemical properties (Color and Chemical properties)and the experimental the therapeutic efficacy of various medicines wasstandardized. Some deviations were found which could be due to theeffect of variable environmental factors influencing the chemicalconstituents of the medicines.

Thus, the method will help to know the therapeutic efficacy of themedicines under study. Hence, the proposed method will become a novelvisual proof for the understanding the therapeutic efficacy of themedicine reported or new, single or formulated.

The analysis of the images was done using software developed for thispurpose. The details of the software is given in the release notes andFIG. 115

Step 4:

Interpretation of the Data

The fingerprints generated are analyzed for their chemical andtherapeutic properties. The basic features in a fingerprint are found tobe

1) the zone of the polarity in which the constituents have eluted; and

2) the conjugative properties of the individual constituents present.

The polarity of the column is fixed. It is a normal phase or a reversedphase stationary phase. In the normal phase column, stationary phasewill be polar and in a reverse phase column, the stationary phase willbe non-polar. The extent of polarity of the stationary phase varies frombrand to brand even in same type of reverse phase or normal phasecolumn. The polarity of the stationary phase will be controlled usingthe polarity of the mobile phase, additives like buffers and pH. Whenthe polarity of the mobile phase is varied constantly in the increasedor decreased order, on a reverse phase column, the constituents presentin the sample will elute in the same order, i.e., the high polarconstituents will be eluted by the high polar mobile phase, the mediumphase mobile phase will elute the medium polar constituents and thenon-polar constituents will be eluted by the non polar or low polarmobile phase. The most preferred pattern is to change the polarity ofthe mobile phase either increased or decreased order of polarity suchthat no constituent of any polarity will be left un-eluted from thecolumn thus achieving total elution. Thus by controlling the polarity ofthe stationary phase, polarity of the mobile phase will be managed tobring a required effect on the polarity of the constituents to achieveseparation of required order of elution.

The order and properties of polarity and elution in the case of normalphase columns are applicable as in the case of reverse phase column butreverse to the reverse phase column. The non-polar constituents willelute first, followed by polar constituents, based on the order ofpolarity of the mobile phase used for elution.

Thus in the present elution also the elution of the constituents iscontrolled and driven in the required pattern by controlling thepolarity of the mobile phase and the order of changing it in an orderlyway.

Mostly the elution of the samples were done from high polarity mobilephase to low polarity mobile phase. Thus in the finger prints theconstituents present in the first zone (Zone-1) will be of high polar innature. The same pattern applies to the other zones, the medium polarconstituents eluted in the medium polar zone (Zone-2) and the low ornon-polar constituents eluted in the non-polar zone (Zone-3). Thispattern reverse when a normal phase column is used due to its elutionproperty as described above.

Most of the high polar molecules will be highly reactive chemically,thus biologically. When they enter the first part of the digestivesystem mouth, they will immediately start acting on the biologicalsystem and the enzymes present there. Then the constituents will enterthe stomach and intestine where they will under go different changes(Post assimilation effects, Vipaka in Ayurveda) due to the digestivejuices and their enzymes present in the part. In the process ofabsorption the moment the molecules of high activity (high polar) willimmediately start interacting with the biological system and show theirtherapeutic properties. This can be compared that in Ayurveda, theintestinal part of the human body is classified as Pitta zone, where thehigh polar molecules are playing a major role. The heat causingmechanism will play an important role in the diseases and biologicalmechanisms related to. It indirectly indicates the molecules of highreactive, the high polar molecules.

After the absorption, the blood with all the absorbed constituents willcarry them to heart and the parts related to it. Then the blood will besent to different parts of the body. In Ayurveda, the upper portion ofthe human body is defined as the Kapha zone, where the cold mechanismwill be playing an important role. Thus, the molecules of medium polarmolecules will play an important role in the mechanisms related to thiszone.

The low polar and non-polar constituents will be able to enter to thehuman body only through blood transfer, Thus the body organs where themechanism of availability of the chemical constituents is only by bloodwill be coming in the last category of the polarity. The non-polar oils,fats and other such molecules and mechanisms in the human body areclassified as Vata disorders and all such disorders are cure using thesame type of materials.

The low and non-polar constituents will be eluting in the last zone ofthe fingerprint. Thus, this zone (ZONE-3) is considered as Vala zone.Thus the basic humors of the molecules are able to be identified as pertheir polarity which facilitates to know on what disorder (dosha) it isgoing to act upon. Thus, the present method is useful for thetherapeutic standardization of the medicines.

The image was divided in to three zones on X and Y-axis. The conjugativeproperty (Absorption of a particular wavelength of radiation) is takenon Y-axis and polarity is taken on the X-axis as the elution of theconstituents is controlled using the polarity of the mobile phasecomposition. Now as reported in literature the Y axis is scaled as per,the therapeutic efficacy based on wavelength (color). The entire imageis divided in to six chambers where in the chemical constituents have aspecific conjugative and polarity property. This in turn is proportionalto the therapeutic efficacy of the constituents in the chamber. Thus,when a medicine is fingerprinted, based on the color represented for theabsorption of a specific wavelength and having a specific polarity, thetotal colors in that zone is calculated and interpreted for thetherapeutic efficacy of the constituents present in it. Thus, theholistic therapeutic standardization and chemical standardization isachieved using this method. A schematic representation is given in table15, showing the relation of conjugation and polarity to therapeuticefficacy of the different constituents present in a medicine.

When the 3-D chromatograms of the medicine will be analyzed using allits 3 dimensional properties of the said image. If the 3-D chromatogramis considered as a cap with a hood the matching of the entire cap 3dimensionally, with another sample of different qualitative andquantitative properties, the extent it matched will be presented as ananalytical report qualitatively and quantitatively. Here the hood of theCap is compared to the peak of the molecule at a particular wavelength.A sample with more number will like a cap with many hoods. Thus thematching of the three dimensional coordinates will provide a foolproofmethod of comparison and analysis. The coordinate it matched will givequalitative and the extent it matched will give the quantitative data ofthe sample understudy. This is made possible by special softwareprepared for this purpose. This becomes an ultimate method of qualitycontrol.

But any method without quantification will be of no use. Hence, thetotal colors of the constituents in the image of a particular zone areconsidered as a representation of the amount of the polar constituentspresent in the medicine. Thus the total constituents present in theZone-1 Pitta zone, Zone-2 Kapha zone, Zone-3 Vata zone are present inthe form of a Pie diagram which represents the ratio of the efficacy ofthe medicine on each of the disorder. Thus, medicines havingconstituents in the order of 50:20:30 will be medicines of Tridoshaharaof the order of 50%: 20%: 30%. This was done using the softwaredeveloped. Thus, the therapeutic efficacy is standardizedquantitatively. The increase or decrease of any one or two of the otherdoshas can be done by formulating medicine by adding other medicines andprepare a suitable formulation needed to cure a specific individual.

The chemical standardization was done using the software by quantifyingthe individual constituents based on the colors denoting theconcentrations of the in-gradients. The range of the wavelength that amolecule absorbed denotes the conjugative properties.

As described in the traditional standardization methods the colors ofthe medicines were standardized based on their colors and theirtherapeutic efficacy. It applies even in the case of any molecules. TheTable 8 of colors and their efficacy will explain how colors were usedto standardize the efficacy of the medicines. The colors of themolecules can be understood by their absorptive properties of theradiation of the Uv-Vis range of radiation. In the Table 10 of colorsand the relation with wavelengths, the colors of the medicines and theircharacteristic wavelengths are given. Based on the structure, functionalgroups, conjugation, and the extent of unsaturation will influence thewavelength of absorption (absorbance maxima) of the molecule. The morethe molecule is conjugated the longer the wavelength of absorption willbe. Hence, the UV-VIS absorbance of any molecule is widely used in thequalitative and quantitative properties of the constituents.

The colors and the therapeutic efficacious of various medicines weregiven in the ancient literature. The colors of the molecules are due toa specific chemical nature of the molecule. The colors of the flameswere used for the quality control of metals and related products, whichinvolves the basic spectrophotometric principles. Thus, study andunderstanding of the interaction of the electromagnetic radiation willbe useful to study the chemical nature and thus the therapeutic efficacyof the medicines. The same principle has been used in the presentspectrophotometric method of fingerprinting and standardization. Inother terms an existing concept has been presented in the form of anovel analytical method, removing the error of human factor. All themedicines for which fingerprints developed were given in table oftherapeutic efficacy of the medicines were given in table 14. Thetechnical details of the software are given in the release notes of thesoftware.

Release Notes for the Software Proposed

I) System Requirements (Minimum)

-   a. Processor: Pentium II or higher-   b. OS: Windows 95, Widows 98, Win NT 4.0 and Linux-   c. RAM: 64 MB or higher-   d. Monitor: 14″ Color Monitor (1024×768) or higher-   e. Software: Java Development Kit (JDK 1.2.X)    II) The Operational Mechanism of the Software:

The various operational mechanisms are described below: operationalsequences with various functionality are shown in FIG. 115.

Title of the Software: RAINBOW (An Image analysis software forchromatographic fingerprints)

This software is developed for the chromatographic fingerprints andmicroscopic images.

-   1. It is GUI (Graphical User Interface) based software.-   2. The software is designed to analyze any kind of image    particularly for the analysis of chromatographic fingerprints.-   3. The reports are given in form of graphs.-   4. Life Cycle    a. In put: Image    b. Processing:-   Analysis involves-   Extracting Colors (Standard 7 colors and some of their different    shades)-   Resizing, Deviding in to 3 zones at 20 minutes interval-   Graphing (Bar and Pie graphs)-   Bar-coding-   Standards Followed for extracting the colors:

The software extracts eight colors viz. Red, Green, Blue, Yellow, Cyan,Magenta and Orange.

Any color is not absolute. It is mixture of the following shades of thecolors present before and after it, they vary between a range of values.The range for the colors used to identify as the colors given above therespective values are taken from the international standard 256-colorscale. The values used in the present software are: Red Blue Green ForRed color 200-255  0-64  0-64 and 192-200  0-64  0-32 For Green color 0-64  0-48 200-255  0-65  0-64  65-191 For Blue Color  0-96 200-255 0-191

Similarly, other colors were taken as standards for the extraction ofcolors. (These standards are exclusive for the present softwarerequirements and are modifiable if required)

While image is analyzed, the software reads the image pixel by pixel andreads and extracts the color according to the color standardsdesignated, stores and transfers them for further display as bar graphs.

c. Output:

Reporting

-   1. By Graphs-   2. By saving data like images, graphs, deviding the image display    into three zones-   3. By displaying the ‘X’ (Retention time or Pixel value of the    image), Y (Wave length or absorbance of the images of contour and    3-D chromatograms respectively), R (Red color), G (Green color) and    B (Blue color) coordinates.-   4. By transferring these values to an in-built bar-coding software    to generate a bar code.    d. User Interaction:

User is allowed to interact with the product in various ways.

-   1. Inputing the desired image (one or more)-   2. Resizing the image to desired size and analyzing it.-   3. Saving the image, resized image, and graphs to it.-   4. Printing the image, resized image, and graphs to it.    III) Technical Features of the Software-   1. It is software entitled ‘Rainbow’-   2. A software with a facility of opening chromatographic fingerprint    images in different Formats (extensions) like BMP, JPEG, TIF, GIF    from the file folders and analyze it for different colors present in    the image with single pixel sensitivity.-   3. A software with a facility of display of the pixel information in    the form of 1.a graph having a scale of X (0-(min. time scale) and Y    (200-800 nm) coordinates and 2. a Pie diagram with individual values    of each peak (Automatic and Manual) in two separate columns beside    the graph.-   4. Software with a facility of printing all the data generated after    analysis using PRINT Icon.-   5. A software with a facility of changing the page setup for    printing using PAGE SETUP Icon-   6. A software with a facility of selecting a part of the image and    analyze using RESIZE Icon.-   7. A software with a facility of opening any number of image    analysis windows for different images, and display of status in    WINDOW icon.-   8. A software with a facility of dividing the image in to three    Zones at 20 min interval, using ZONE icon.-   9. A software with a facility of inverting the selected image using    INVERT icon.-   10. A software with a facility of switching over to Notepad, Word    pad and MS Word, using EDITOR icon.-   11. A software with a facility of operational information about    various features of the Software using, the HELP icon.-   12. Software with a facility of saving the data generated using SAVE    AS icon as. JEPG file format.    IV) Installation Instructions for the Software:    a. Installation Procedure of Java 1.2.x Soft Ware Platform on Which    the Present Software Works.-   Explore the Java CD-ROM-   Double click on the jdk1.2.0/jdk1.2.1/jdk1.2.2 setup icon-   The setup will extract the files and conforms from the user whether    to load the software in the system.-   On click, ‘yes’ it asks for the directory into which it should    install the files.-   By default, c:\jdk1.2 directory will be shown.-   If you want to install in “d” drive, change the directory and    Install the software.-   Once the installation is completed go to c: and open the file named    ‘autoexec.bat’.-   Give the following path in the autoexec.bat file.-   Open auto exe. Bat and write as follows-   Set path=d:\jdk1.2\bin: % path %-   Setup class path=d:\jdk1.2\lib\classes.jar; % classpath %    Reboot and Use    b. Installation of Image Analyzer Software Proposed-   1. Copy the folder of the image analyzer software from the CD on to    the system in the desired directory.-   2. Explore the batch file form the software folder into which image    analyzer software was copied.-   3. Right click on it and click on ‘send to desktop as short cut’-   4. A ‘MS dos’ iconic short cut appears on the desktop. Right click    the icon and go to properties, select program tab and check on the    ‘close on exit check box’, convert window status ‘To minimized’.-   5. Apply and close.-   6. Now the image analyzer software is ready for use. Double click on    the image analyzer Icon and it starts working.-   7. In the opening window, a box with ‘CSIR’ will open where in the    pass word ‘dvk’ should be typed.-   8. Click the arrow mark (hand) on the right down corner of the    opening image to open the Software.-   9. Open the directory of the images of contour images without scale    and select the image to be analyzed. The image will be shown on the    image window.-   10. Click on the RED analytical window marked with red boarder. A    PIE diagram will be displayed along with a chromatogram with    retention time on X-axis and nanometers on the Y-axis.-   11. For constituents of lesser concentration click on the Green,    Yellow and Orange colors. The Other colors are mostly the base line    or less than that hence can be ignored.-   12. The details of using the other features of the software are    given in the help menu of the software including the various    features and applications of the software.    V. Known Bugs:

Not found

VI. Abbreviations Used:

-   a. JDK: Java Development Kit-   b. Con: Contour Chromatogram-   c. 3-D: 3-Dimensional Chromatogram-   d. WOS: Without Scale-   e. X: Represents the Retention Time of the chromatogram-   f. Y: Represents the absorbance in the 3-D chromatogram and wave    length range in contour chromatogram-   g. R: Intensity of red color at a particular pixel position-   h. G: Intensity of green color at a particular pixel position-   i. B: Intensity of blue color at a particular pixel position    VII. Meaning of the Various Icons and functions

a. PRINT icon will facilitate in printing all the data generated afteranalysis.

b. PAGE SETUP icon will facilitate in changing the page setup forprinting.

c. RESIZE icon will facilitate the selection of a part of the image andanalyze the selected part of the image.

d. WINDOW icon will facilitate in opening any number of image analysiswindows for different images, and display of status in

e. ZONE icon will facilitate in dividing the image in to three Zones at20-min interval.

f. INVERT icon will facilitate in inverting the selected image.

g. EDITOR icon will facilitate in switching over to Notepad, Word padand MS Word.

h. HELP icon will facilitate in the operational information aboutvarious features of the Software usage.

i. SAVE AS icon will facilitate in saving the data generated in a *.JPEGfile format.

VIII. Constraints:

a) The present software works only for contour chromatograms withoutscale.

b) The scale on X axis is in 1-(of “minutes” representing retention timeof the contour chromatogram

c) The scale on Y-axis is 200-800 nm representing the range ofwavelength under which the analysis is conducted.

d) The Image developed has to be resized using imaging software to matchthe run time and wavelength range on X and Y-axis.

e) The images after analysis will be saved only in JPEG format withoutscale.

f) The clip images should be stored with an extension of theco-ordinates i.e. X1 and Y2.

Main Advantages of the Present Invention are:

-   1. The contour chromatogram of the medicine becomes a Fingerprint of    it. Because it contains the UV-Vis spectrum band with concentration    of the ingredients along with the polarity of the molecule. The    fingerprints developed for a same medicine extracted under different    pH value helps to understand the drug release in the intestine    system at different pH values thus facilitating the pharmacodynamics    of the medicines under study.-   2. The spectral bands of all the constituents are given in a single    picture assessing the medicine about its therapeutic properties and    nature, very easy.-   3. The 3-D Chromatogram becomes a photo of all the UV-spectra of all    wavelengths of each constituent in a single picture indicating the    chemical (conjugative and polarity) property of the molecule eluted.-   4. A database of the fingerprints of various herbal medicines    available in the country useful for Quality control, Forensic and    customs departments to control the use and misuse of the herbal    medicines at the public interest.-   5. The database also gives information about the medicinal value of    the various medicinal plants of the country (therapeutically    classified) in the country. and the role of the ecological factors    on the chemical constituents of the same plant available in various    tropical Zones of the country. This facilitates to select a plant    for collection of the herbal medicines suitable to be used for the    therapeutic usage for a medicinal professional or an herbal trader.-   6. Analysis of the fingerprints using this software gives role of    ecological factors on various herbal medicines available in the    country and it is useful for Quality Control, Forensic and Customs    Departments to control use and misuse of the herbal medicines at the    national interest-   7. The analysis of the fingerprints is useful to understand the    therapeutic efficacy of the medicines using the physico-chemical    properties of the medicines as reported in the ancient literature.-   8. The analysis also gives information about the medicinal value of    the various medicinal plants in the country and the role of the    ecological factors on the chemical ingredients of the same medicine    available in various parts of the country.-   9. The therapeutic and ethano-botanical classification of the    fingerprints helps to bring some generalizations useful for the    doctors and researchers for a complete understanding of the    traditional medicines by analyzing the fingerprints.-   10. By bar coding the image properties the medicines/plant    extracts/plants are saved from piracy as the facility to create a    barcode using the properties of the image through note pad facility.-   11. The barcodes are utilized in all commercial transactions of    modern ERP and CRM applications.    Application Utilities of the Present Invention International

It is useful for any country for fingerprinting and patenting thetraditional medicines of that country. Because the finger print of asingle medicinal plant is not similar to a finger print of the sameplant in another place or country due to the variations in its chemicalprofile. The variations in chemical profile is due to the influence ofEcological factors like the tropical region variations, soil, waterquality and the genotypic and phenotypic variations factors on thechemistry of the plant.

This method helps the country to fulfill one of the regulations made byWHO, for the member countries to standardize methods for the utility andquality control of herbal medicines and their regulation.

National

This is useful as a tool to prevent international piracy of traditionalmedicines by Passing a Law that the “The Medicinal plants for which thefinger prints are developed are national property”. If a medicine isapplied for any type of patent, in any place of the world and if thefinger print, tallies with the finger print of the medicine available inthe challenging country, the patent could be objected.

Strategic

Bar coding the fingerprints of the medicines helps in the authenticregulation and protection of the medicinal plants.

By converting the barcode of the fingerprint of a medicinal plant into amachine-readable language, commercial and regulatory work becomes easy.

Fingerprints of the medicines helps the Food and Drug controllers,Customs and Central Excise departments to regulate, check the use,misuse and pilferage of the herbal medicines inside the country andwhile allowing importing of such medicines.

Industrial

A fingerprint developed for a medicine or a formulation helps theindustry to protect their process technology by comparing thefingerprint of the same medicine of other brands. Thus, it helps toimplement the patent law more efficiently.

The fingerprint helps to monitor how the medicines are changing itsmedicinal properties by the addition of another medicine at differentstages of process of preparing a formulation.

Industry can use the database of fingerprints developed for all thenative plants available in the country, for their selection of the placeof collection of a medicine. It helps the industry, which part of thecountry, and in which season is suitable for the collection of herbalmedicines, as the ecological factor changes the therapeutic efficacy ofthe medicines.

Scientific

This method helps the researchers to understand the traditionalformulations prepared. It also helps to monitor a new formulation underpreparation.

It helps to know how new molecules are formed when a complicatedtraditional formulation is prepared.

The fingerprints developed for a same medicine extracted under differentpH value helps to understand the drug release in the intestine system atdifferent pH values of an individual.

The contour chromratogram of the medicine becomes a Fingerprint of it.Because it contains the UV-Vis spectrum band with concentration of theingredients along with the polarity of the molecule.

The spectral bands of all the constituents are given in a singlepicture, assess the medicine about its therapeutic properties andnature, very easily.

The 3-D Chromatogram becomes a photo of all the UV-spectra of allwavelengths of each constituent in a single picture indicating thechemical (conjugative and polarity) property of the molecule eluted

The database also gives information about the medicinal value of thevarious medicinal plants of the country (therapeutically classified) inthe country and the role of the ecological factors on the chemicalconstituents of the same plant available in various tropical Zones ofthe country. This facilitates to select a plant for collection of theherbal medicines suitable to be used for the therapeutic usage for amedical professional or an herbal trader.

The therapeutic and ethano-botanical classification of the fingerprintshelps to bring some generalizations useful for the doctors andresearchers for complete understanding of the traditional medicines byanalyzing the fingerprints.

The present method facilitates to prepare chromatographic fingerprinting of herbal medicines and formulations, which is useful for manyquality control and regulation purposes.

The present method facilitates chemical standardization, (qualitativeand quantitative) of the said medicines by providing the conjugative andpolarity properties of the individual molecules present in the medicinesor any organic or organo-metallic compound which has UV-VIS absorptiveproperty. This kind of analysis is of much use in the chromatographicanalysis of Herbal medicines of Single and Formulations, where the useof external or internal standards are practically not possible.

The invention facilitates to study, understand and monitor thetherapeutic efficacy of the said medicine under study. It helps tounderstand the therapeutic actions and properties of traditionalmedicines reported in the ancient literature and confirm the same in theform of a reproducible analytical data. Thus, it provides therapeuticstandardization of the medicines under study. It shows the polarityzones like Polar, Medium polar and Non polar molecules present in thesample, thus facilitating to understand the efficacy of the medicine asa whole.

This method facilitates to re-standardize the reported medicines to thepresent therapeutic needs. It helps to monitor and study formation ofnew organic and organo-metallic molecules, which has UV-VIS absorptiveproperty, in the process of preparing a reported or a new formulation.This also helps to standardize the process technology of preparing areported or new formulation by monitoring the constituents and theirchanging chemical and therapeutic properties.

It facilitates to generate a barcode by an in built bar coding software,wherein the X retention time, Y wavelength, R number of red pixels, Gnumber of green pixels and B number of blue pixels are the coordinatesgiven by the present software. Some examples of the barcode forchromatograms are given. The invention also facilitates bar coding oneor more of the constituents present in the fingerprint thus facilitatingthe commercial transactions easy by ENTERPRISE RESOURCE PLANNING (ERP)and CUSTOMER RELATIONSHIP MANAGEMENT (CRM) applications. A database thusprepared helps the regulatory authorities to monitor the movement of thesaid medicines inside or from outside the country, from production tothe consumer. The database of the barcodes thus prepared, becomes theresource for the ERP vending machines or of any of such kind. Themachine will display all details of the medicines like company, itschemical fingerprint, and the therapeutic efficacy of the medicines thesaid medicine. This makes the identification of the medicines moreauthentic than the present.

A data base of the fingerprints thus generated using this method helpsto bring many generalizations of the therapeutic efficacy of aparticular therapeutic class of plants. Thus, one can under stand why aparticular plant is added in that class. This is explained in the Table14 enclosed.

The fingerprints printed on the label of the medicine helps the doctorsto understand the therapeutic efficacy of the medicine just before useand confirm the quality control of the medicines for every batch.

The Image analysis of Chromatographic Finger Print Images (ContourChromatograms) of various Medicines of any philosophies (Single andFormulations) developed, are useful for many purposes as described invarious steps of this application.

Social

This is useful to know for a consumer, the therapeutic efficacy ofmedicines single or formulated claimed on the label and confirms tocontain the same.

This helps the consumer-act to monitor the quality control of herbalmedicines sold in the market and protect the interests of the consumer.

Adulteration

The Image analysis of Chromatographic Finger Print Images (ContourChromatbgrams) of various Medicines of any philosophies (Single andFormulations) developed is useful for detection of any adulteration ofthe medicines. TABLE 1 Table of Different Philosophies And VariousTerminology Used in Medicine Sl TRI PANCHA SAPTA TRI No PHILOSOPHY DOSHA(Hara) BHUTA PROPERTIES DHATU MALAS 1 Ayurveda Vata, 1. Prithivi 1.Rasa-Taste-6 1. Rasa 1. Purisha (Shown elaborately Pitta, 2. Ap 2.Guna-Property-Basically 20 2. Rakta 2. Mutra in seperate table) Kapha.3. Teja 3. Veerya-Potency-2 3. Mamsa 3. Sweda Binary: 4. Vayu 4.Vipaka-Metabolite-3 4. Medas Prakriti-Purusha 5. Akasha 5.Prabhava-Specific properties- 5. Asthi Innumerable 6. Majja 6.karma-Action 7. Shukra 2 Siddha Pitta, 1. Mann(Prithivi) 1. Rasam 1.Rattam (Blood) 1. Malam Binary: Kapha, 2. Neer (Ap) 2. Gunam 2. Sadhai(Muscle) 2. Mutram Prakriti-Purusha Vata 3. Thee (Agni) 3. Veeryam 3.Kozhuppu (Medas) 3. Vervai 4. Vayu (Vayu) 4. Vipakam 4. Elumbu (Bone) 5.Akasa (Akasha) 5. Vindhu, Karu (Sperm, Ovum) 3 Chinese 1. Yang 1.Wood 1. Sour 1. Sinuses Information Binary: Yin-Yang 2. Yin 2. Fire 2.Bitter 2. Blood vessels not 3. Earth 3. Sweet 3. Muscles available* 4.Metal 4. Shark 4. hair 5. Water 5. Salty 5. Bones 4 Tibetan Nes Pas 1.Sa (Prithvi) Information not available* Lus Zuns Bdun Dri Ma 1. Mkhris(Pitta) 2. Chu (Ala) 1. Dans Ma (Rasa) 1. Bsan 2. Bad-Kan (Kapha) 3. Me(Agni) 2. Khrg (Rakta) 2. Gcin 3. Rlun (Vata, 4. Rluin (Vayu) 3. Sa(Mamsa) 3. Rnul Vayu) 4. Tsil (Medas) 5. Rvs (Asthi) 6. Rkan (Majja) 7.Khu Ba (Shukra) 5 Unani Akhalat Arkan 1. Garm (Hot) 1Primary: Blood,Phlegm, Bile, 1. Bole Binary: 1. Damvi (Blood) 1. Aag (Fire) 2. Khush(Dry) Saudai 2. Baraj Normal-Abnormal 2. Balgam (Phlegm) 2. Hawa (Air)3. Sard (Cold) Secondary: 3. Paseena 3. Safravi (Bile) 3. Pani (Warter)4. Motadil (Neutral) 1. Mahsoora(Intravascular) 4. Saudai (Vata) 4.Earth (Mitti) 2. Talliya (Pericellular) 3. Qureeba (Intercellular) 4.Munviya (Cellular) Body organs Simple, Compound 6 Greek 1. YellowBile 1. Water 1. Hot 1. Unctuous Contrary medicine 2. Black Bile 2.Earth 2. Dry 2. Rough 3. Phlegm 3. Fire 3. Wet 3. Hot 4. Blood 4. Air 4.Cold 4. Cold

TABLE 2 Relation Of Humors, Properties, And Different Parts Of The HumanBody - An Ayurvedic Approach Approach RELATION PANCHA EFFECT ON ON BHUTADOSHAS VIPAKA (PHYSI- (DECREASING (POST TRI CAL MAHABHUTA THE DOSHA)ASSIM- Sl. DOSHA TRI PROPER- SAPTA CHEMICAL RELATIONS DUE TO ILATIVE No(Hara) MALAS TIES) DHATUS PROPERTIES WITH DHATUS DHATUS RELATION ON GUNAEFFECT) 1. Vata, 1. Purisha 1. Prithivi 1. Rasa 1. Rasa a. Prithivi + Apa. Pitta Vata Hara a. Guru, Sheeta, snigdha a. Madhura 2. Pitta, 2.Mutra 2. Ap 2. Rakta (Shadruchi's) b. Agni + Prithive b. Vata Hara b.Ushna, Laghu, snigdha b. Amla 3. Kapha 3. Sweda 3. Teja 3. Mamsa a.Madhura c. Jala + Agni c. Vata Hara c. Ushna, Laghu, Snigdha c. Madhura4. Vayu 4. Medas b. Amla d. Aksha + Vayu d. Kapha Hara d. Ushna, Laghu,Ruksha d. Katu 5. Akasha 5. Asthi c. Lavana Eagni + Vayu e. Kapha Pittae. Sheeta, Laghu, Ruksha c. Katu 6. Majja d. Katu f. Prithive + VayuHara f. Sheeta, Guru, Ruksha f. Katu 7. Shukra e. Tikta f. Kapha Pittaf. Kashaya Hara 2. Guna-: Broadly classified into 3 groups 1. Vaisheshik2. Samanya 3. Atma Mostly used are: Guru (Heavy) Laghu (Light) Sheeta(Cold) Ushna (Hot) Snigdha (Soft, Lubricated, Supple) Rooksha (Dry)Manda (Slow) Teekshna (Sharp) 3. Veerya-2 4. Vipaka-3 5. Prabhava-innumerableThe properties of the materials and disease were correlated to select amedicine - The therapeutic standardization

TABLE 3 Table Showing Division in terms of the Macrocosm in Chinesemedicine Sl After No. Elements Seasons Color Taste influence Development1 Wood Spring Blue Sour Wind Birth 2 Fire Summer Red Bitter Heat Growth3 Earth Late Yellow Sweet Humidity Puberty Summer 4 Metal Autumn WhiteShark Drought Maturity 5 Water Winter Black Salty Cold SenilityBasis of color for therapeutic standardization

TABLE 4 Table Showing Division In Terms Of the Microcosm in ChineseMedicine SL Sense Structural No. Elements organs Fus Tsangs Emotionaleluents 1 Wood Eye Bile Liver Anger Sinuses 2 Fire Tongue Small HeartJoy Blood intestine Vessels 3 Earth Mouth Stomach Spleen Anxity Muscle 4Metal Nose Large Lung Sadness Hair intestine 5 Water Ear Bladder KidneFear Bones

TABLE 5 Table Showing the Relation of Five Natural Elements and TheirRelation, in Chinese Medicine Element Element Yin & Yang 1^(st) Heavenengendered Water 2^(nd) Energy engendered Fire corresponds to the Heart& Small intestine 3^(rd) Heaven engendered Wood 4^(th) Energy engenderedMetal corresponds to the Lung & Large intestine 5^(th) Heaven engenderedEarth 6^(th) Energy completed Water corresponds to the Kidney & Bladder7^(th) Heaven completed Fire 8^(th) Energy completed Wood corresponds tothe Liver & Gall bladder 9^(th) Heaven completed Metal 10^(th)  Energycompleted Earth corresponds to the Spleen & Stomach

TABLE 6 Table Showing the Meaning of Yin and Yang Used In ChineseMedicine Yin Yang (The shady side of the hill) (The sunny side of thehill) Light Dark Night Day Damp Dry Cold Hot Water Fire Evil Good UglyBeautiful Vice Virtue Poverty Healthy Sadness Joy Confusion OrderPunishment Reward Disease Health Positive Negative Female Male Bad GoodWife Husband

TABLE 7 Table Showing The Basis Of Color For The TherapeuticStandardization Of Medicines Color White Color Medicines Yellow ColorMedicines Red Color Medicines Black Color Medicines Name in SancritShukla Varga Peeta Varga Rakta varga Krishna Varga Sloka

Name of the Sudha Chuna (Lime) Kusumba Pushpa Dadima(Punica granatum)Kadali (Musa paradisiaca) medicines/ Kachhapa Prista (Shell of Kimshuka(Butea Palasha(Butea monosperma) Karavellika(Momordia charantia)materials tortoise) monosperma) Laksha(Laccifera lacca) Triphala(Threemyribalans) Shankha (Conch) Haridra (Curcuma longa) Bandhuka Neelika(Indigofera Tinctoria) Shukti (Shell of Pearl) Patanga (Caesalpiniasappan) Haridra (Curcuma longa) Nala (Cymbophogan species) Varatika(Small shells) Madayantika(Lasonia Kusumba Pushpa Panka (Lotus)Brushtashma (Incinerated Stone) Inermis) Manjista (Rubia cordifolia)Kaseesa(Fe₂S₃) Sarkara (Sugar candy) *Rasamava *Rajanighantu Balamra(Unripen Mango) *Rajanighantu by Vaidya *Rasendra Chudamani NarahariThe names of the medicines were given as sloka

TABLE 8 The Effect Of Different Colors On Different Diseases Cool effectNeutral Creat heat Violet Indigo Blue Green Yellow Orange Red 1. Bones& 1. E-N-T problems 1. Whooping cough, 1. Heart problems 1. Alldisorders 1. Long term 1. Anaemia Bone marrow, 2. Facial paralysis 2.Throat problems, 2. Low and high of digestion Asthma 2. Disability 2.Tumour, 3. Diseases of the lungs, 3. Fever, B. P 2. Spleen, liver 2.bronchitis 3. Sluggishness 3. Baldness, 4. Asthma, 4. Typhoid, 3. Skinproblems problems 3. Swelling in 4. Cold 4. Cataract, 5. T. B, 5.Small-pox, 4. Cancer 3. Diabetis trachea 5. Paralysis 5. Blindness 6.Less digestive power, 6. Measles, 5. Influenza 4. Leprosy etc., 4. Gout6. White spots 7. Problems of nervous 7. Ulcers in mouth 6. Syphilis 5.Swelling 7. Arthrites systems, 8. Cholera 7. pain in the eyes 6. Kidney8. T. B etc., 8. Convulsion, 9. Swelling in the etc., 7. Mental 9.Lunacy brain nervousness 10. Problems of 8. Epilepsy nerves etc., 11.Insomnia, 12. Mental depression 13. Problems of semen discharge 14.Burns, Bleeding from nose etc.,The role of colors and their influence on different body parts, Thebasis of color is used for selecting a medicine. For eg. A plant withIndigo flowers will cure ENT problems

TABLE 9 Properties of the SIX Tastes (Rasas in Ayurveda) and theirproperties and efficacy Predominent Examples Taste element Effects onDosha Dietary Item Drug Sweet (Madhur) Earth + Water Kapha ↑ Sugar,Banana, Jack fruit, Glycerrhiza Glabra., Vata and Pitta ↓ Raisins, Milk,Coconuts, Asparagus Racemoses, Jaggary Gold Sour (Amla) Earth + FirePitta and Kapha ↑ Tamarind, Buttermilk, Curds, Embalika officinalis Vata↓ Raw mango Salty (Lavana) Water + Fire Pitta and Kapha ↑ Salt Rock saltVata ↓ Pungent (Katu) Air + Fire Pitta and Vata ↑ Asoefetida, Pepper,Chilli, Dry Piper longum Kapha ↓ processed Zinger Bitter (Tikta) Space +Wind Vata ↑ Bitter gourd Azadiracta indica, Pitta and Kapha ↓ Swertiachiraita, Tinospora Cordifolia Astringent (Kashaya) Wind + Earth Vata ↑Honey Terminalia chebula, Pitta and Kapha ↓ Treminalia Bellerica,Pearls, CoralsThis table shows how tastes and medicines are related to vitiation ofdiseases.

TABLE 10 Table or Colors and The Relation with Wavelengths. WavelengthColor Color observed (Transmitted) Nm (Absorbed from white light) orComplementary Hue* <380 Ultraviolet Yellowish green 380-435 VioletYellow 435-480 Blue Orange 480-490 Greenish blue Red 490-560 Bluishgreen Purple 500-560 Green Violet 560-580 Yellowish green Blue 580-595Yellow Greenish blue 595-650 Orange Bluish green 650-780 Red >780 Near -Infra red*The constituents having these colors will absorb at the respectivewavelengths given The materials or medicines will show colors based onthe absorption of a particular color from the range of colors in thewhite light falling on them. They will express the resultant color afterabsorption.

TABLE 12 Comparison table of existing techniques Sl No ReportedTechnique Method Merits Demerits 1. TLC Fingure Printing 1. Simple, 1.Inferior compared to Closed chromatography Open 2. Less time consuming,2. Incomplete Separations leading to ambiguous Chromatography 3. Lessoperational Costs separations 3. Unreliable fingure printing due toinfluence of variations in analytical conditions 4. Needs the support ofother costlier analytical instruments like LC-MS, NMR and IR with outwhich the data is incomplete. 2. HPTLC Fingure Printing 1. Simple 1.Inferior compared to Closed chromatography. Open 2. Less time consuming2. Incomplete Separations leading to ambiguous Chromatography 3. Lessoperational Costs separations. 3. Unreliable fingure printing due toinfluence of variations in analytical conditions. 4. Needs the supportof other costilier analytical instruments like LC-MS, NMR and IR without which the data is incomplete. 5. High instrumental cost 3. HPLC 1.Chromatogram 1. Better separations 1. Needs the support of othercostilier analytical Closed at a specified 2. facility to change theinstruments like LC-MS, NMR and IR with out Chromatography wave lengthpolarity of the mobile phase to which the data is incomplete. (Superiorthan open elute all molecules of entire 2. High instrumental costchromatography) range of polarity. 3. High operational cost 4.PROPOSED 1. Chromatogram 1. Better separations 1. High instrumental costMETHOD indicates the 2. facility to change the (almost equal or lessthan a HPTLC instrument) HPLC entire range of polarity of the mobilephase to 2. High operational cost Closed wave length. elute allmolecules of entire Chromatography 2. Utilisation of range of polarity.OPERATIONAL LIMITATIONS OF THE (Superior than open CONTOUR 3. Facilityto measure the PRESENT COMPUTER BASED METHOD chromatography)CHROMATOGRAMS absorbance of the molecules at 1. This computer basedmethod works only for for the various wave lengths of the contourchromatograms with out scale. analysis of the entire range of 200-800nm. 2. The Image developed has to be resized using organic and This Willnot leave any imaging software to match the run time and Organo-metallicmolecule UNSEEN or wavelength range on X and Y-axis. molecules 3.UNIDENTIFIED. 3. The images after analysis will be saved only inUtilisation of 3-D 4. Facilitates to prepare the JPEG format with outscale which occupies less CHROMATOGRAMS “Chromatographic Finger memoryof the system. for the Prints” of the native medicinal 4. The clipimages should be stored with an analysis of the plants of a country asextension of the co-ordinates ie., X1 and Y2. organic and suggested byWHO. How ever they can be eliminated by the addition Organo-metallic 5.Facilitates to understand the of more software features moleculestherapeutic efficacy of the medicines by studying the conjugative andpolarity properties of the constituents separated by this method. 6.Facilitates to understand the therapeutic efficacy of a particulartherapeutic class of plant (just like the method used in theidentification of the personality of a culprit, as used in the FINGERPRINT SOFTWARE used by the Forensic departments.

TABLE 13 PARAMETERS USED FOR FINGER PRINTING OF MEDICINES 3-DPARAMETERS* FIG BOTANICAL NAME OF VERNACULAR ELEVATION ROTATION NO. THEPLANT NAME PART USED DEGREES DEGREES 29 ABEL MOSCHUS KASTURI BENDA WHOLEPLANT 20 15 MOSCHATUS MEDICUM 30 ACACIA SUMA SWETHAKHADIRA BARK 15 65 31ACALYPHA INDICA KUPPINTA LEAF LETS 10 60 32 ADHATODA VASAKA VASA LEAVES25 45 33 ADIANTUM CAUDATUM MAYURASHIKHI LEAVES 20 40 34 AILANTHUSEXCELSA ARALU STEM BARK 10 65 35 ACORUS CALAMUS VACHA RHIZOME 10 130 36ALLIUM PORUM MAHALASUNA LASSAN, BIG 20 130 SINGLE CLOVES 37 ALLIUMSATIVAM LASUNA LASSAN, 20 130 SMALL CLOVES 38 ALPINIA GALANGA GREATERRHIZOME 20 75 GALANGA. 39 ALPINIA OFFICINARUM LESSER GALANGA RHIZOME 1575 40 ALPINIA SPECIOSA LIGHTER RHIZOME 10 60 GALANGA 41 ARECA CATECHUBEETLE NUT UN PROCESSED 15 40 FRUIT NUT 42 ARECA CATECHU BEETLE NUT MILK15 40 PROCESSED NUTS 43 ARECA KATEEH RAKTHA STEM BARK 15 65 KHADIRA 44ARNICA ARNICA MOTHER 10 55 TINCTURE OF WHOLE PLANT 45 BACOPA MONNERIBRAHMI WHOLE HERB 15 45 46 BERBERIS ARISTATA DARUHARIDRA STEM AND 15 170BARK 47 BORRHIEVIA DIFFUSA PUNARNAVA WHOLE PLANT 15 55 48 CAPSCICUMANNUM L MIRCH BIG, RIPED 10 70 FRUIT 49 CAPSCICUM ANNUM L MIRCH BIGUNRIPED 10 70 FRUIT 50 CAPSCICUM ANNUM L MIRCH SMALL, 10 70 UNRIPED,FRUIT 51 COSCINIUM LATA DARVI STEM BARK 15 125 FENESTRATIUM 52COCCINIDIUM GRANDIS DONDA ROOT AND 25 30 LEAF 53 DACTLYLACTINIUM GRASSLEAF 25 40 AEGYPTIUM(ERECT) 54 DACTLYLACTINIUM GRASS LEAF 25 40AEGYPTIUM(PROSTRATE) 55 DIRISTACHIS TUMMA LEAF AND 20 15 CINERARIA BARK56 EMBLICA OFFICINALIS AMALAKI FRUIT EPICARP 5 50 57 FACE PACK BRAND 1FORMULATION 20 25 58 FACE PACK BRAND 2 FORMULATION 20 25 59 GLYCERRHZIAGLABRA YASHTI MADHU ROOT BARK 15 130 60 GLYCERRHZIA GLABRA YASHTI MADHUPOWDER OF 15 130 WHOLE PALNT 61 GYMNEMA PODAPATRI WHOLE PLANT 25 15SYLVESTRAE 62 HOLLERONA KUTAJA STEM BARK 10 60 ANTIDYSENTRICA 63 INNULARECEMOSA PUSHKARAMULA ROOT 5 45 64 MICHELLIA CHAMPAKA MANU SAMPENGAFLOWER 20 40 65 MORINGA OLIFERA MUNAGA LEAF 25 40 66 MYRICA CEREFERA BAYBERRY MOMEOPATHIC 20 35 MOTHER TINCTURE 67 NAHI AXILLAE NAHI WHOLE PLANT10 130 68 OROXYLUM INDICUM SYONAKA STEM BARK 10 170 69 OCIMUM SANCTUMRAMA TULASI LEAF 15 130 70 PLUCHEA PATRA RASNA LEAF 10 65 LANCEOLATA 71PICRORRHIZA KURROH KATUKI ROHINI STEM BARK 15 125 72 PIPER BEETLE BEETLELEAF 25 160 73 PSORALIA CORILIFOLIA BAKUCHI SEEDS 25 60 74 RAPHANUSSATIVUS MULLANGI, WHITE LEAF 15 25 75 RICINUS CUMMUNIS ERANDA MULA ROOT10 135 76 RUBIA CORDIFOLIA MANJISTA STEM AND 10 40 ROOT 77 SAUSSREALAPPA KUSHTA ROOT 5 80 78 SPHERANTHUS INDICUS MUNDI WHOLE HERB 15 70 79SYMPLOCUS LODHRA STEM BARK 15 65 RACEMOSUS 80 TERMINALIA CHEBULAHARITAKI FRUIT 10 40 81 TERMINALIA VIBHITAKI FRUIT 20 35 BELLERICA 82TRIGONELLA FAENUMG. MENTHI WHOLE PLANT 15 160 83 TRIBULUS GOSHURA STEMAND 25 45 TERRESTRIAS ROOT 84 TYLOPHORA LEAVES 10 65 ASTHMATICA 85VIBURNUM MOTHER MOTHER 20 15 TINCTURE TINCTURE OF HOMOEO MEDICINE 86WITHINIA SOMNIFERA ASWAGANDHA ROOT 5 50 87 ZINZIBER OFFICINALIS SHUNTIPROCESSED 15 130 ZINGER, RHIZOME 88 AVIPATTAKARA AYURVEDA POWDER 25 60CHURNA FORMULATION 89 KAMADUGA SIDDHA POWDER 10 25 FORMULATION 90KUMARAYASAVA AYURVEDIC LIQUID 10 35 MEDDICINE BY FERMENTATION PROCESS 91MAHALAKSHMI VILAS SIDDHA POWDER 20 35 RAS FORMULATION 92 SUVARNAYOGARAJA SIDDHA POWDER 10 40 GUGGULU FORMULATIONALL THE OTHER PARAMETERS OF RANGE OF WAVE LENGTH, ABSORBANCE SCALE ANDRETENTION TIMES ARE SHOWN IN INDIVIDUAL FIGURE

TABLE 14 MEDICINES USED FOR FINGER PRINTING DOSHA HARA (Disorder onwhich PACIFIES) BOTANICAL NAME OF THE PLANT VERNACULAR NAME EFFICACY*PITTA KAPHA VATA PART USED PITTA HARA AVIPATTAKARA AYURVEDA Laxative, ↓POWDER CHURNA FORMULATION Peptic ulcer, Piles ACALIPHA INDICA HARITAHepatoprotectitive, ↓ LEAF MANJARI Skin diseases, Gyneac disordersANANDABHAIRAVI HERBOMINERAL Pitta jwara FORMULATION AROGYA VARDHINIHERBOMINERAL Liver disorders, ↓ FORMULATION Skin disorders BHUMYAMALAKIPHYLLANTHUS Jaundice ↓ WHOLE HERB URINARIA KAMADUGA FORMULATION Pepticulcer ↓ FORMULATION KUMARAYASAVA FERMENTATION Gyenic disorders, ↓ LIQUIDPROCESS Jaundice SARACA INDICA ASHOKA Gyeneac disorders ↓ STEM BARKSURYAVARTI HERBOMINERAL Head ache ↓ FORMULATION KAPHA HARA AILANTHUSEXCELSA ARALU Digestive disorders ↓ ASPARAGUS SAFED MUSALI Aphrodisiac,↓ ROOT ADESCENDENTUM impotency ADHATODA VASICA VASA Respiratory ↓ ROOTdisorders ADIANTUM CAUDATUM MAYURASHIKHI Piles ↓ WHOLE HERB CoughDiarrhoea ALLIUM SATIVAM LASUNA Swasa ↓ SMALL CLOVES ALLIUM PORUMMAHALASUNA Swasa ↓ BIG SINGLE CLOVE ACACIA SUMA SWETHA Diabetis ↓ STEMBARK KHADIRA CAPSCICUM ANNUM L KATUVEERA Digestive disorders ↓ BIGUNRIPED, FRUIT COCCINIDIUM GRANDIS BIMBI Emitic ↓ CHOPACHINYADI HERBALVenerial, ↓ POWDER CHURNAM FORMULATION Skin diseases GLYCERRHZIA GLABRAYASHTI MADHU Panduroga, ↓ ROOT, BARK Rasayana HIBISCUS ABEL LATA KASTURISiheshma roga, ↓ WHOLE PLANT MOSCHUS Prameha, urinary WITH FLOWERSblader &kidney AND SEEDS disorders INNULA RECEMOSA PUSHKARAMULA Kasa,swasa, jaundice, ↓ ROOT diabetis KRIMIKUTARA RAS HERBO-MINERAL Worminfestation ↓ FORMULATION OCIMUM SANCTUM RAMA TULASI Cough, ↓ LEAF FeverRAPHANUS SATIVUS MULKA, WHITE Diabetics, cough, g. ↓ LEAF i.tractdisorders, neutraceuticals SAUSSREA LAPPA KUSHTA Respiratory ↓ ROOTdisorders SHILAJIT (H) HERBOMINERAL Diabetis, ↓ BITUMINOUS Renal stonesSHILAJIT (G) HERBOMINERAL Diabetis, ↓ BITUMINOUS Renal stones TYLOPHORAAJADWESHI Asthama LEAF ASTHAMATICA VATA HARA ALPINIA GALANGA GREATERRheumatic ↓ RHIZOME GALANGA, disorders ALPINIA LESSER Rheumatic ↓RHIZOME OFFICINARUM GALANGA disorders ALPINIA SPECIOSA LIGHTER Rheumatic↓ RHIZOME GALANGA disorders BRIHATVATACHINTAMANI + HERBO-MINERALArthritis ↓ FORMULATION SWARNAMAKSHKAM BORRHIEVIA PUNARNAVA Odema, ↓WHOLE PLANT DIFFUSA Urinary tract, diuretic disorders HUTHASANAHERBO-MINERAL All types of fevers ↓ FORMULATION MAHAYOGARAJA HERBALArthrites ↓ FORMULATION GUGGULU PLUCHEA PATRA RASNA Rheumatic ↓ LEAFLANCEOLATA disorders RICINUS COMMUNIS ERANDA MULA Constipation, ↓ ROOTRheumatoid disorders SUVARNA YOGARAJA SIDDHA Rheumatic diseases ↓ POWDERGUGGULU FORMULATION SHITAMSU RAS HERBO-MINERAL Jwara ↓ FORMULATIONSUVARNA YOGARAJA HERBAL Arthrites ↓ FORMULATION GUGGULU FORMULATION VATAGAJANKUSA RAS HERBO-MINERAL Sciatica ↓ FORMULATION PITTA KAPHA HARAACACIA CATECHU RAKTHA Skin diseases diabetis ↓ ↓ STEM BARK, KHADIRAEXUDATE AILANTHUS EXCELSA ARALU Digestive disorders ↓ ↓ BARK ARECACATECHU KRAMUKA Diabetis, ↓ ↓ MILK PROCESSED Skin disorders NUTAZARIDICTA INDICA NIMBA Skin diseases, ↓ ↓ TENDER LEAVES Infectiveconditions BERBERIS ARISTATA DARUHARIDRA Obesity ↓ ↓ ROOT BARK Skindisorders CITRULLUS INDRAVARUNI Purgative, juandice, ↓ ↓ HOMOEO MOTHERCOLOSYNTHIS aborttificient TINCTURE CURCUMA LONGA TURMERIC Worminfestation, ↓ ↓ COMMERCIAL dysentry, diarrhoea, skin POWDER-1disorders, wounds CURCUMA LONGA TURMERIC Worm infestation, ↓ ↓COMMERCIAL dysentry, diarrhoea, skin POWDER-2 disorders, wounds CURCUMALONGA TURMERIC Worm infestation ↓ ↓ COMMERCIAL dysentry, diarrhoea, skinPOWDER-3 disorders Wounds COSCINIUM LATA DARVI Diabetis ↓ ↓ STEMFENESTRATIUM Obesity, Skin disorders CURCUMA LONGA HARIDRA Skin, ↓ ↓ RAWRHIZOME Allegic, Diabetic DACTYLACTINIUM GRASS Diuretic, improves ↓ ↓WHOLE PLANT AEGIPTIUM complexion LEAVES (PROSTRATE AND ERECT) EUGENIAJAMBU Vomiting, diabetic, ↓ ↓ FRUIT JAMBOLONA Dysentry SIZYGIUM CUMINIHOLLERNA KUTAJA Diarrhoea, ↓ ↓ STEM BARK ANTIDYSENTRICA Haemorroids FROMANDHRA PREADESH HOLLERNA KUTAJA Diarrhoea, ↓ ↓ STEM BARK ANTIDYSENTRICAAll gi tract disorders FROM KERALA RUBIA CORDIFOLIA MANJISTA Skindisorders ↓ ↓ STEM, ROOT Leukemia Blood purifier PSORALIACORYLIFOLIABAKUCHI Leucoderma, ↓ ↓ SEEDS, SEED OIL Skin diseases PICRORRHIZA KATUKALaxative ↓ ↓ ROOT KURROA ROHINI Liver disorders TRIGONELLA METHIKADiabetis, colic ↓ ↓ WHOLE HERB FENUM G. SYMPLOCOS LODHRA Bleeding ↓ ↓BARK RACEMOSA disorders, diarrhoea, dysentry SPERANTHUS INDICUS MUNDIKrimihara, ↓ ↓ WHOLE HERB Migrain, vrishya, lymphatic disorders KAPHAVATA HARA ACORUS CALAMUS VACHA Medhya ↓ ↓ RHIZOME Speeh disordersALOEVERA KUMARI Gyneac disorders, ↓ ↓ LEAF, LEAF JUICE hepatomegaly,spleeno megaly, burns, uterine disorders AGNITUNDINA HERBAL Indigestion,↑ FORMULATION FORMULATION Skin disorders MICHELIA CHAMPAKA CHAMPAKACosmetic, ↓ ↓ FLOWER Skin diseases MORINGA OLEIFERA SIGRU Abcess, oedema↓ ↓ LEAF PIPER BEETLE NAGA VALLI Kasa, swasa ↑ ↓ LEAVES FROM Digestivedisorders COASTAL ANDHRA PRADESH PIPER BEETLE NAGA VALLI Kasa, swasa ↑ ↓LEAVES Digestive disorders CULCATTA TRIKATU CHURNA-1 HERBAL Indigestion↓ ↓ FORMULATION FORMULATION TRIKATU CHURNA SP-2 HERBAL Indigestion ↓ ↓FORMULATION FORMULATION TRIKATU CHURNA GH-3 HERBAL Indigestion ↓ ↓FORMULATION FORMULATION TRIBULUS TERRESTRIAS GOKSHURA Urinary disorders,↓ ↓ STEM AND Oedema ROOT TYLOPHORA AJADWESHI Diabetis, asthama ↓ ↓LEAVES ASTHMATICA PITTA VATA HARA ACACIA SUMA SWETHA Prameha ↓ ↓ STEMBARK KHADIRA ANANDABHAIRAVI HERBO- ↓ ↓ FORMULATION MINERAL BACOPAMONNERI BRAHMI Medhya, skin disorders ↑ WHOLE HERB DICROSTACHYS CINERAVEERATARU, Hridya, ↓ ↓ LEAF AND BARK TUMMA Obescity ENICOSTEMMA NAHIMalaria ↓ ↓ WHOLE HERB AXILLAE KANCHANARA HERBAL Inflammatory conditions↓ ↓ FORMULATION GUGGULU FORMULATION OROXYLUM INDICUM SYONAKA Odema ↓ ↓STEM BARK Digestive disorders TRI DOSHA HARA ASPARAGUS SWETHAAphrodisiac ↓ ↓ ↓ ROOT ABSCENDENSES MUSALI CAPSCICUM ANNUM L MIRCH, G.Itract disorders ↓ ↓ ↓ BIG, RIPED FRUIT KATIVEERA CURCULIGO KALI MUSALIAphrodisiac ↓ ↓ ↓ ROOT ORCHIOIDES EMBLICA OFFICINALIS AMALAKI Hridya, ↓↓ ↓ FRUIT EPICARP Rasaya, na Neutraceutical KARPURADI RAS HAEMORRHOIDSDiarrhoea ↓ ↓ ↓ FORMULATION MAHALAKSHMI VILAS HERBO-MINERAL All types offevers ↓ ↓ ↓ FORMULATION RAS ONION BIG PALANDU Haemorrhoids ↓ ↓ ↓ BULBONION SMALL PALANDU Haemorrhoids ↓ ↓ ↓ BULB TERMINALIA CHEBULA HARITAKILaxative ↓ ↓ ↓ FRUIT Rasayana TERMINALIA VIBHITAKI Kasa, swasa, ↓ ↓ ↓FRUIT BELLERICA Skin diseases, urinary calliculus WITHINIA PUB.ASWAGANDHA General debility, ↓ ↓ ↓ ROOT RED SEEDS rejuvenation PLANTROOT ADULTERATIONS FACE PACK (G) Good by efficacy FORMULATION FACEPACK(B) Absence of some FORMULATION important constituents like kushta(sausserea lappa) and manjista (rubia cordifolia), making theformulation less effective HERBAL HEAD BATH No foaming agents foundFORMULATION POWDER (G) HERBAL HEAD BATH Adulterated with FORMULATIONPOWDER (B) foaming agents HOMOEO MEDICINES ARNICA Pain reliever, afterHOMOEO MOTHER effects of injury TINCTURE MYRICA CEREFERA BAY BERRYHepatoprotective HOMOEO MOTHER TINCTURE BRAND-1 MYRICA CEREFERA BAYBERRY Hepatoprotective HOMOEO MOTHER TINCTURE BRAND-2 VIBURNUM MOTHERGyneacological disorders HOMOEO TINCTURE MEDICINE ISOLATED COMPOUNDSVITEX NEGUNDO SINGLE Hepatoprotective A FLAVONOID COMPOUND 7-HYDROXYQUERCETIN AZAMALYCIN SINGLE SINGLE ISOLATED COMPOUND COMPOUND A DIABETICHERBAL Prameha ADULTERATED WITH MEDICINE A DIURETIC ALLOPATHIC AMIDECONSTITUENT BERBERIS ARISTATA SINGLE BERBARIN STD ISOLATED (FLUKA)STANDARD COMPOUND↓: INDICATES DECREASE OF DISORDER↑: INDICATES INCREASE OF DISORDER*The therapeutic efficacys reported are as per the information availablewith the author, many more may be reported elsewhere.

TABLE 15 Names of the medicines shown as thumbnails of fingerprints NAMEOF THE SAMPLE PART USED Allopathic medicines Vitamin-B complexCommercial brand Analgin Commercial brand Atenolol Commercial brandBromoflexin Commercial brand Citerizine Commercial brand FurazolidineCommercial brand Ibuprofen-Paracetamol Commercial brand ParacetamolCommercial brand Herbal Cosmetic samples Face pack (Poor by efficacy)Commercial brand Face pack (Good by efficacy) Commercial brand Head bathpowder (Poor by efficacy) Commercial brand Head bath powder (Good byefficacy) Commercial brand Herbal Formulations Agnitundina Commercialbrand Anandabhairavi Commercial brand Arogyavardhani Commercial brandBrihatvatachintamani + Commercial brand Swarnamakshakam ChopachinyadiChurnam Commercial brand Commercial Turmeric Brand 1 Commercial brandCommercial Turmeric Brand 2 Commercial brand Commercial Turmeric Brand 3Commercial brand Huthasana Commercial brand Homoeo medicines ArnicaMother Tincture Calendula Mother Tincture Colosynthis Mother TinctureIsolated compounds 7-Hydroxy Quercetin Isolated from Vitex negundo Aflavonoid Isolated from Vitex negundo Azamalycin Standard DexamethasoneStandard Single medicines Alovera Leaf Acalypha Indica Leaf Embalikaofficinalis Fruit Aralu Bark Areca Catachu Seeds Aswagandha Red fruitsRoots Aswagandha White fruits Roots Beetle leaf Andhra pradesh LeafBeetle leaf Culcutta Leaf Coscinium Stem Bark Dactlylactenium aegyptium(Erect) Leaves Dacylactinium aegyptium (Prostrate) Leaves DaruharidraStam Bark Haridra Raw Rhizome Kalajamun Fruit Onion Bulb Kutaja StemBark Raktakhadira Heart wood Shilajit Processed Bituminous Source 1Shilajit Processed Bituminous Source 2 Brahmi Leaf

TABLE 16 The Division of the Fingerprint in to Therapeutic Zone based onthe conjugation and polarity

Based on the color reported, the entire image is divided in to 3 Zoneson X-axis and 3 Zones on Y-axis. X axis shows the POLARITY SCALE due tothe mobile phase composition. Y-axis shows CONJUGATION (due to UV-VISabsorbance. When the molecule is more conjugated it absorbs at higherwavelengths (800 nm).# Thus constituents present in the respective zones will act as shown inthe figure in the respective therapeutic zones will be providingrespective therapeutic efficacy. Quantification of these constituentswas done using the UV-VIS absorptive property which is directlyproportional to the quantity of the constituent.

1. A method for detection and identification of principles from extractsof plants or animal, natural or synthetic sources, using chromatographicfinger printing techniques, said method comprising the steps of: i)extracting the organic or organo-metallic molecules using a suitablesolvent; ii) subjecting the extract obtained in step (i) to theseparation analysis, using High Pressure Liquid Chromatographytechniques; iii. generating contour and 3D chromatograms of theingredients eluted based on the pH and polarity; iv. converting the 3-Dand contour chromatogram obtained into a colored image, analyzing thecolored image for its individual colors using the coordinates denotingall its 3-dimensional properties of the said image by using an inbuiltsoftware; v. denoting the concentrations of the various constituentseluted with time; vi. generating a chromatogram based on color analyzed,having peaks at various retention times along with conjugativeproperties of the molecules; vii. identifying the compounds in the saidingredients by the UV-Vis absorptive properties of the variousconstituents in the image; viii. identifying, determining andclassifying the compounds eluted as polar, medium polar and less ornon-polar based on the polarity and conjugative properties; ix.generating a barcode for a selected peak using X axis as retention time,Y axis as wavelength, R as number of red pixels, G as number of greenpixels and B as number of blue pixels; and x. generating a database offingerprints and barcodes and identifying the respective compounds inthe samples.
 2. A method as claimed in claim 1 wherein, the solventswith different polarities are selected based on the hydrophilic andhydrophobic nature of the sample under study, ethyl alcohol is used forstandardization of medicines.
 3. A method as claimed in claim 1 wherein,the fingerprints are developed for the same medicine extracted underdifferent pH ranges.
 4. A method as claimed in claim 1 wherein, the HPLCapparatus used is selected from any commercially available HPLCapparatus with the Photo Diode Array detector, preferably with agradient or ternary system of pumps.
 5. A method as claimed in claim 1wherein, the pH and polarity of the mobile phase is controlled byvarying the ratio of the mixture 0 to 100% of an aqueous solvent, wateror a buffer at a required pH by using a salt (like Potassium Di-Hydrogenorthophosphate or Di potassium hydrogen orthophosphate and phosphoricacid maintaining the required pH) with a non-aqueous solvent.
 6. Amethod as claimed in claim 1 wherein, the non-aqueous, organic andaqueous, water or buffer at a known pH are the solvents used in step 1(iii) and are selected based on the range of polarity.
 7. A method asclaimed in claim 1 wherein, converting the contour chromatograms into acolored image comprising the conjugative and polarity properties of theconstituents of the medicine under study.
 8. A method as claimed inclaim 1 wherein, the therapeutic efficacy of a medicine (single orformulated) is assessed using the quality of the constituents present ina particular polarity and UV-Vis absorptive zone.
 9. A method as claimedin claim 1 wherein, the software generates a barcode for a selected peakor peaks or image using the X axis as Retention Time, Y axis asWavelength, R as number of Red Pixels, G as number of Green Pixels and Bas number of Blue Pixels as the coordinates, provided by the software,which makes the product propriety for an industry.
 10. A method asclaimed in claim 1 wherein the software used is called Rainbow havingthe following features: (a) a software with a facility of openingchromatographic fingerprint images in different Formats (extensions)like .BMP, JPEG, TIF, GIF from the file folders and analyze it fordifferent colors present in the image with single pixel sensitivity; (b)a software with a facility of display of the pixel information in theform of 1.a graph having a scale of X (0-(min. time scale) and Y(200-800 nm) coordinates and
 2. a Pie diagram with individual values ofeach peak (Automatic and Manual) in two separate columns beside thegraph; (c) software with a facility of printing all the data generatedafter analysis using PRINT Icon; (d) a software with a facility ofchanging the page setup for printing using PAGE SETUP Icon; (e) asoftware with a facility of selecting a part of the image and analyzeusing RESIZE Icon; (f) a software with a facility of opening any numberof image analysis windows for different images, and display of status inWINDOW icon; (g) a software with a facility of dividing the image in tothree Zones at 20 min interval, using ZONE icon; (h) a software with afacility of inverting the selected image using INVERT icon; (i) asoftware with a facility of switching over to Notepad, Word pad and MSWord, using EDITOR icon; (j) a software with a facility of operationalinformation about various features of the Software using, the HELP icon;and (k) software with a facility of saving the data generated using SAVEAS icon as. JEPG file format.
 11. A software based data processing of 3D chromatograms and color contour image of an ingredient, saidprocessing comprising: a. analyzing (extracting colors) the coloredcontour image based on the selection of various colors (with standardsmentioned in release notes, life cycle, processing) denoting theconcentrations of the various constituents eluted with time, andpolarity based on retention time; b. analyzing the 3-D chromatograms ofthe medicine using all its 3 dimensional properties of the image; c.generating a chromatogram having peaks at various retention times alongwith conjugative properties of the molecules eluted with time in aspecified order of polarity; d. identifying the compounds in the saidmolecules by the UV-Vis absorptive properties of the variousconstituents in the image; e. correlating the reported biological,therapeutic activity of the of various constituents present in themedicines understudy based on the polarity and the conjugativeproperties of the molecules by dividing the fingerprint into therapeuticzones on X and Y axis; f. generating a barcode for a selected peak(s)using the image coordinates viz., X for retention time, Y forwavelength, R for number of red pixels, G for number of green pixels andB for number of blue pixels, provided by the proposed software; g.generating a database of fingerprints and barcodes for the samples,facilitating all kinds of database utilities like Enterprise ResourcePlanning (ERP) and Customer Resource Management (CRM) applications; andh. generating a database of the ‘display widows’ for all the samples tobe used by the ENTERPRISE RESOURCE PLANNING (ERP) and CUSTOMERRELATIONSHIP MANAGEMENT (CRM) type of business applications.
 12. Amethod as claimed in claim 11 wherein, the solvents used for extractionis selected based on the polarity, hydrophilic and hydrophobic nature ofthe constituents, sample and its constituents under study.
 13. A methodas claimed as claimed in claim 11 wherein, the HPLC apparatus used isselected from any commercially available HPLC apparatus with the PhotoDiode Array detector, preferably with a gradient or ternary system ofpumps.
 14. A method as claimed in claim 11 wherein, the polarity of themobile phase of a non-aqueous and an aqueous solvent of a specific pH iscontrolled by varying the ratio of the mobile phase from 0% to 100% ofan aqueous solvents like water or a buffer of a known pH, along with anon-aqueous solvent or vice-versa.
 15. A method as claimed in claim 11wherein, on analysis of 3-D and contour chromatograms using new softwarecalled Rainbow, that gives a chromatogram with retention time andwavelength on its X and Y-axis.
 16. A method as claimed in claim 11,wherein, on analysis of 3-D and contour chromatograms using new softwarewhich gives a data having indicated the vitiation of doshasquantitatively in percentage ratio.
 17. A method as claimed in claim 11wherein, a single solvent ethanol is used for extraction of theconstituents; same analytical conditions and instrumental parameterswere used for all samples to bring the therapeutic generalizations. Thetherapeutic standardization is thus achieved.
 18. A method as claimed inclaim 11 wherein the software used is called Rainbow having thefollowing features: i. a software with a facility of openingchromatographic fingerprint images in different Formats (extensions)like .BMP, JPEG, TIF, GIF from the file folders and analyze it fordifferent colors present in the image with single pixel sensitivity; ii.a software with a facility of display of the pixel information in theform of 1.a graph having a scale of X (0-(min. time scale) and Y(200-800 nm) coordinates and
 2. a Pie diagram with individual values ofeach peak (Automatic and Manual) in two separate columns beside thegraph; iii. software with a facility of printing all the data generatedafter analysis using PRINT Icon; iv. a software with a facility ofchanging the page setup for printing using PAGE SETUP Icon; v. asoftware with a facility of selecting a part of the image and analyzeusing RESIZE Icon; vi. a software with a facility of opening any numberof image analysis windows for different images, and display of status inWINDOW icon; vii. a software with a facility of dividing the image in tothree Zones at 20 min interval, using ZONE icon; viii. a software with afacility of inverting the selected image using INVERT icon; ix. asoftware with a facility of switching over to Notepad, Word pad and MSWord, using EDITOR icon; x. a software with a facility of operationalinformation about various features of the Software using, the HELP icon;and xi. software with a facility of saving the data generated using SAVEAS icon as. JEPG file format.
 19. A computational method ofchromatographic finger printing, chemical and therapeuticstandardization and bar coding of organic and organo-metallic moleculesfrom a plant, animal or a naturally available or man made materials usedas medicines, said method comprising a) selection of medicines andextraction of the constituents, b) separation of the constituents intoindividual constituents, generating and converting the 3-D and contourchromatograms into fingerprints, c) analyzing the fingerprints using thesoftware developed, and d) interpretation the data.
 20. A method asclaimed in claim 19 wherein, it provides chemical analysis of theconstituents present in the medicine under study and their conjugativeand polarity properties indicating the therapeutic efficacy as per thetraditional concepts of the medicine using the new software developed.21. A method as claimed in claim 19 wherein, it provides a novel conceptof chromatographic finger printing of herbal medicines which is usefulfor the quick identification of the actual profile of the compoundspresent in the medicine under use along with their therapeutic efficacyof the constituents.
 22. A method as claimed in claim 19 wherein, itprovides a novel chromatographic finger printing of herbal medicines andformulations using the contour and 3-D chromatograms of the herbalmedicines and formulations is proposed and they are developed on a PhotoDiode Array Detector (PDA) of a High Pressure Liquid Chromatograph. Thisdelineates the data of the spectral properties of the constituentspresent in the herbal medicines presented in a specific order ofpolarity under similar experimental analytical conditions.
 23. A methodas claimed in claim 19 wherein, said method provides UV-Visible spectraof the compounds having displayed the conjugative and polarityproperties of the molecules and the concentration of the individualconcentrations of the molecules along with the polarity of themolecules.
 24. A method as claimed in claim 19 wherein, said methodprovides UV-VIS spectra of all the constituents shown in a single image“The Chromatographic Fingerprint”, the said fingerprint becomes the blueprint of the constituents present in an herbal medicine or formulationfor an assay and quick identification of the medicine understudy.
 25. Amethod as claimed in claim 24 wherein, same standard analyticalparameters like extraction with same solvent ethyl alcohol, same runtime 0-60 min, same mobile phase acetonitrile along with phosphatebuffer having a pH in the range of 5.5-7.5, and a same UV-Visible Rangeof 200-800 nm for fingerprinting and chemical and therapeuticstandardization.
 26. A method as claimed in claim 24 wherein,fingerprinting is used for the study of adulterated, substituted,contradictual and commercial food and drug samples and to identify thepure and impure.
 27. A method as claimed in claim 24 wherein,fingerprinting method is used for identifying the chemical constituentspresent in it for the purpose of process standardization, qualitycontrol activities and therapeutic standardization of Allopathic,Ayurvedic, Homoeo, Siddha, Unani, Chinese, Tibetan, Kampo (Japanese)medicines.
 28. A method as claimed in claim 24 wherein, fingerprintingmethod is used for the study of variation of chemical constituents dueto various ecological factors, geological factors, genotypic andphenotypic variations (in plants) in naturally occurring samples and toidentify and standardize the chemical constituents in them.
 29. A methodas claimed in claim 24 wherein, fingerprinting is used for the study ofchemical constituents in synthetically prepared samples and to identifyand standardize the chemical constituents in them for chemical andtherapeutic standardization which ever is applicable.
 30. A method asclaimed in claim 24 wherein, fingerprinting is used for the study ofchemical constituents in herbal products of single medicine samples andto identify the chemical constituents in them for chemical andtherapeutic standardization.
 31. A method as claimed in claim 24wherein, fingerprinting is used for the study of chemical constituentsin herbal products of formulated medicine samples and to identify thechemical constituents in them for chemical and therapeuticstandardization.
 32. A method as claimed in claim 24 wherein,fingerprinting is used for the study of variation of chemicalconstituents in biological samples and to identify and standardize thechemical constituents in them.
 33. A method as claimed in claim 24wherein, fingerprinting is used for the study of variation of chemicalconstituents in different brands of products of single and formulatedfood and medicine samples and to identify the chemical constituents inthem for chemical and therapeutic standardization.
 34. A method asclaimed in claim 24 wherein, preparation of a database of a large numbersamples gives many generalizations of the therapeutic efficacy of aparticular group of plants, classified as a group for a particulardisease or therapeutic classification.
 35. A method as claimed in claim24 wherein, fingerprinting of medicines facilitates to categorize andquantify the constituents of a medicine based on polarity andconjugation from 3-D and contour chromatograms and assess thetherapeutic efficacy of the medicine on which humors it is going to act(vitiate).
 36. A method as claimed in claim 24 wherein, fingerprintingenables to understand and standardize the Physico-Chemical properties ofthe medicines like color for the use of therapeutic standardization ofmedicines and humors using conjugative and polarity properties given inthe chromatographic fingerprints.
 37. A method as claimed in claim 24wherein, the fingerprinting method enables to understand and standardizethe Physico-Chemical properties of the medicines like Tastes (Rasa) likeSour, Salty, Pungent, Bitter, Astringent (Amla, Lavana, Katu, Tikta,Kashaya as described in Ayurveda) used for therapeutic standardizationusing conjugative and polarity properties shown in the chromatographicfingerprints.
 38. A method as claimed in claim 24 wherein, thefingerprinting method enables to understand and standardize thePhysico-Chemical properties of the medicines like Property, Potency,Metabolite, Specific properties like Chirality of the molecules (Guna,Veerya Vipaka, Prabhava) used for the therapeutic standardization usingconjugative and polarity properties of the individual constituents andthe whole medicine shown in the chromatographic fingerprints.
 39. Amethod as claimed in claim 24 wherein, the fingerprinting method enablesto understand and standardize the Physico-Chemical properties (Gunas) ofthe medicines like Cold, Hot, Slow in action, Sharp in action, Heavy,Light, Soft Lubricated Supple, Dry (Sheeta, Ushna, Manda, Teekshna,Guru, Laghu, Snigdha, Rooksha as described in Ayurveda) used for thetherapeutic standardization using conjugative and polarity properties ofthe medicines shown in chromatographic fingerprints.
 40. A softwarebased data processor of 3 D chromatograms and color contour image of aningredient, said processor comprising computing means and capable of: i)an analyzer (extracting colors) for analyzing the colored contour imagebased on the selection of various colors (with standards mentioned inrelease notes, life cycle, processing) denoting the concentrations ofthe various constituents eluted with time, and polarity based onretention time; ii) an analyzer for analyzing the 3-D chromatograms ofthe medicine using all its 3 dimensional properties of the image; iii) ameans for generating a chromatogram having peaks at various retentiontimes along with conjugative properties of the molecules eluted withtime in a specified order of polarity; iv) an identifier for identifyingthe compounds in the said molecules by the UV-Vis absorptive propertiesof the various constituents in the image; v) a means for correlating thereported biological, therapeutic activity of the of various constituentspresent in the medicines understudy based on the polarity and theconjugative properties of the molecules by dividing the fingerprint intotherapeutic zones on X and Y axis; vi) a means for generating a barcodefor a selected peak(s) using the image coordinates viz., X for retentiontime, Y for wavelength, R for number of red pixels, G for number ofgreen pixels and B for number of blue pixels, provided by the proposedsoftware; vii) a means for generating a database of fingerprints andbarcodes for the samples, facilitating all kinds of database utilitieslike Enterprise Resource Planning (ERP) and Customer Resource Management(CRM) applications; and viii) a means for generating a database of the‘display widows’ for all the samples to be used by the ENTERPRISERESOURCE PLANNING (ERP) and CUSTOMER RELATIONSHIP MANAGEMENT (CRM) typeof business applications.
 41. A processor as claimed in claim 40wherein, the solvents used for extraction is selected based on thepolarity, hydrophilic and hydrophobic nature of the constituents, sampleand its constituents under study.
 42. A processor as claimed in claim 40wherein, the HPLC apparatus used is selected from any commerciallyavailable HPLC apparatus with the Photo Diode Array detector, preferablywith a gradient or ternary system of pumps.
 43. A processor as claimedin claim 40 wherein, the polarity of the mobile phase of a non-aqueousand an aqueous solvent of a specific pH is controlled by varying theratio of the mobile phase from 0% to 100% of an aqueous solvents likewater or a buffer of a known pH, along with a non-aqueous solvent orvice-versa.
 44. A processor as claimed in claim 40 wherein, on analysisof 3-D and contour chromatograms using new software entitled “Rainbow”that gives a chromatogram with retention time and wavelength on its Xand Y-axis.
 45. A processor as claimed in claim 40 wherein, on analysisof 3-D and contour chromatograms using new software which gives a datahaving indicated the vitiation of doshas quantitatively in percentageratio.
 46. A processor as claimed in claim 40 wherein, a single solventethanol is used for extraction of the constituents; same analyticalconditions and instrumental parameters were used for all samples tobring the therapeutic generalizations. The therapeutic standardizationis thus achieved.
 47. A processor as claimed in claim 40 wherein, thesoftware Rainbow has the following features: (a) It is software entitled‘Rainbow’; (b) A software with a facility of opening chromatographicfingerprint images in different Formats (extensions) like .BMP, JPEG,TIF, GIF from the file folders and analyze it for different colorspresent in the image with single pixel sensitivity; (c) A software witha facility of display of the pixel information in the form of 1.a graphhaving a scale of X (0-(min. time scale) and Y (200-800 nm) coordinatesand
 2. a Pie diagram with individual values of each peak (Automatic andManual) in two separate columns beside the graph; (d) Software with afacility of printing all the data generated after analysis using PRINTIcon; (e) A software with a facility of changing the page setup forprinting using PAGE SETUP Icon; (f) A software with a facility ofselecting a part of the image and analyze using RESIZE Icon; (g) Asoftware with a facility of opening any number of image analysis windowsfor different images, and display of status in WINDOW icon; (h) Asoftware with a facility of dividing the image in to three Zones at 20min interval, using ZONE icon; (i) A software with a facility ofinverting the selected image using INVERT icon; (j) A software with afacility of switching over to Notepad, Word pad and MS Word, usingEDITOR icon; (k) A software with a facility of operational informationabout various features of the Software using, the HELP icon; and (l)Software with a facility of saving the data generated using SAVE AS iconas JEPG file format.
 48. Use of fingerprints of contour and 3-Dchromatograms of the chemical are the basis for identification ofchemical constituents to limit the scope of the invention.